U.S. patent number 7,931,905 [Application Number 12/469,770] was granted by the patent office on 2011-04-26 for cynomolgus gp80 receptor and uses thereof.
This patent grant is currently assigned to Centocor Ortho Biotech Inc.. Invention is credited to Michael Naso, Ronald Swanson, Bethany Swencki-Underwood.
United States Patent |
7,931,905 |
Naso , et al. |
April 26, 2011 |
Cynomolgus gp80 receptor and uses thereof
Abstract
Isolated polynucleotides encoding cynomolgus monkey gp80,
polypeptides obtainable from expression of these polynucleotides,
compositions, recombinant cells, methods of making and using these
polynucleotides, polypeptides, and compositions are useful in
development of human therapeutics.
Inventors: |
Naso; Michael (Radnor, PA),
Swanson; Ronald (San Diego, CA), Swencki-Underwood;
Bethany (Radnor, PA) |
Assignee: |
Centocor Ortho Biotech Inc.
(Horsham, PA)
|
Family
ID: |
41342402 |
Appl.
No.: |
12/469,770 |
Filed: |
May 21, 2009 |
Prior Publication Data
|
|
|
|
Document
Identifier |
Publication Date |
|
US 20090291455 A1 |
Nov 26, 2009 |
|
Related U.S. Patent Documents
|
|
|
|
|
|
|
Application
Number |
Filing Date |
Patent Number |
Issue Date |
|
|
61055237 |
May 22, 2008 |
|
|
|
|
Current U.S.
Class: |
424/185.1;
424/198.1; 424/139.1; 530/350; 424/143.1 |
Current CPC
Class: |
G01N
33/6863 (20130101); C07K 14/7155 (20130101); G01N
33/5088 (20130101) |
Current International
Class: |
A61K
39/00 (20060101); A61K 39/395 (20060101); C07K
14/00 (20060101) |
References Cited
[Referenced By]
U.S. Patent Documents
|
|
|
5807715 |
September 1998 |
Morrison et al. |
|
Other References
Yamasaki et al., "Cloning and Expression of the Human Interleukin-6
(BSF-2/IFN.beta. 2) Receptor," Science 241: 825-828 (1988). cited
by other .
Novick, et al., "Soluble Cytokine Receptors Are Present in Normal
Human Urine," Journal of Experimental Medicine, 170: 1409-1414
(1989). cited by other .
Yasukawa et al., "Purification and Characterization of Soluble
Human IL-6 Receptor Expressed in CHO Cells," Journal of
Biochemistry, 108: 673-676 (1990). cited by other .
Kohler et al., "Continuous cultures of fused cells secreting
antibody of predefined specificity," Nature, 256: 495-497 (1975).
cited by other .
Queen, et al., "A humanized antibody that binds to the interleukin
2 receptor," Proceedings of the National Academy of Science, USA,
86: 10029-10033 (1989). cited by other .
Hodgson, et al., "Making Monoclonals in Microbes," Bio/Technology,
9: 421-421 (1991). cited by other .
Lonberg, et al., "Antigen-specific human antibodies from mice
comprising four distinct genetic modifications," Nature, 368:
856-859 (1994). cited by other .
Knappik, et al., "Fully Synthetic Human Combinatorial Antibody
Libraries (HuCAL) Based on Modular Consensus Frameworks and CDRs
Randomized with Trinucleotides," Journal of Molecular Biology, 296:
57-86 (2000). cited by other .
Mendez, et al., "Functional transplant of megabase human
immunoglobulin loci recapitulates human antibody response in mice,"
Nature Genetics, 15: 146-156 (1997). cited by other .
Krebs, et al, "High-throughput generation and engineering of
recombinant human antibodies," Journal of Immunological Methods,
254: 67-84 (2001). cited by other .
Fishwild, et al., "High avidity human IgG.kappa. monoclonal
antibodies from a novel strain of minilocus transgenic mice,"
Nature Biotechnology 14, 845-851 (1996). cited by other .
Gentz et al., "Bioassay for trans-activation using purified human
immunodeficiency virus tat-encoded protein: Trans-activation
requires mRNA synthesis," Proceedings of the National Academy of
Science USA, 86: 821-824 (1989). cited by other .
Genbank Accession No. XM 001114404, Jun. 14, 2006. cited by
other.
|
Primary Examiner: Landsman; Robert
Assistant Examiner: Hissong; Bruce D
Attorney, Agent or Firm: Dichter; Eric
Parent Case Text
CROSS-REFERENCE TO RELATED APPLICATIONS
This application claims the benefit of U.S. Provisional Application
Ser. No. 61/055,237, filed 22 May 2008, the entire contents of
which is incorporated herein by reference in its entirety.
Claims
What is claimed:
1. An isolated polypeptide comprising a peptide chain having the
amino acid sequence set forth in SEQ ID NO:2.
2. An isolated polypeptide comprising a peptide chain encoded by
the nucleotide sequence set forth in SEQ ID NO:1.
3. An isolated polypeptide comprising a peptide chain having the
amino acid sequence set forth in SEQ ID NO:6.
4. An isolated polypeptide comprising a peptide chain encoded by
the nucleotide sequence set forth in SEQ ID NO:5.
5. An isolated polypeptide comprising a peptide chain having the
amino acid sequence set forth in SEQ ID NO:10.
6. A peptide chain produced by a method for expressing a peptide
chain comprising the steps of: (a) providing an RNA coding for a
peptide chain having an amino acid sequence selected from the group
consisting of SEQ ID NOS:2, 6, and 10; (b) providing the components
of a cell-free expression system; (c) initiating cell free
expression from the RNA provided; and (d) recovering the peptide
chain.
7. The peptide chain of claim 6, wherein the method further
comprises, after the recovering step, confirming expression of a
peptide chain comprising the amino acid sequence selected from the
group consisting of SEQ ID NOS:2, 6, and 10.
8. The peptide chain of claim 7, wherein the cell-free expression
system is selected from the group consisting of a reticulocyte
lystate-based expression system, a wheat germ extract-based
expression system, and an Escherichia coli extract-based expression
system.
9. A method for assessing the safety of a therapeutic candidate
through the use of a gp80 therapeutic candidate comprising:
a)providing a gp80 therapeutic candidate, a first cynomolgus
monkey, a second cynomolgus monkey, wherein the gp80 therapeutic
candidate is an antibody to cyno gp80, an antibody fragment to cyno
gp80, or a combination of the antibody and the antibody fragment,
and wherein the antibody, antibody fragment, or combination is
directed to the cyno gp80 sequence comprising the amino acid
sequence of a member from the group consisting of SEQ ID NOS: 2, 6,
and 10; b) administering the gp80 therapeutic candidate to the
first cynomolgus monkey; and c) determining whether the first
cynomolgus monkey is presenting a deleterious symptom relative to
the second monkey, wherein presentation of a deleterious symptom by
the first cynomolgus monkey shows the gp80 therapeutic candidate is
unsafe, and lack of presentation of a deleterious symptom by the
first cynomolgus monkey shows the gp80 therapeutic candidate is
safe.
Description
FIELD OF THE INVENTION
The present invention relates to the cynomolgus monkey gp80
receptor and uses thereof.
BACKGROUND OF THE INVENTION
IL-6 is a pleiotropic proinflammatory cytokine produced and
secreted by a wide variety of cell types, most notably antigen
presenting cells, T and B cells. IL-6 is involved in such diverse
activities as B cell growth and differentiation, T cell activation,
hematopoiesis, osteoclast activation, keratinocyte growth, neuronal
growth and hepatocyte activation.
IL-6 plays an important role in B cell abnormalities as
demonstrated in systemic lupus erythematosus, multiple myeloma and
lymphoproliferative disorders. Similarly, IL-6 is also implicated
in the pathogenesis of autoimmune and inflammatory diseases, such
as rheumatoid arthritis and osteoarthritis. Evidence also suggests
an association between IL-6 and chronic obstructive pulmonary
disease and insulin resistance in type 2 diabetes. IL-6 has both
pro-inflammatory and anti-inflammatory effects in the immune
system, indicating that this cytokine likely plays a central role
in regulating the physiological response to disease. Therefore,
targeting IL-6 can potentially provide therapeutic benefit in a
variety of disease areas.
An increase in the production of IL-6 has been observed in a number
of diseases including: Alzheimer's disease, autoimmune diseases,
such as rheumatoid arthritis, inflammation, myocardial infarction,
Paget's disease, osteoporosis, solid tumors (renal cell carcinoma),
prostatic and bladder cancers, neurological cancers, and B-cell
malignancies (e.g., Casteleman's disease, certain lymphomas,
chronic lymphocytic leukemia, and multiple myeloma). Research has
indicated that IL-6 is linked to the pathogenesis of many of these
diseases, particularly, cancer and, therefore, blocking IL-6 should
translate into clinical benefits.
IL-6 induces signaling through a cell surface heterodimeric
receptor complex composed of a ligand binding subunit (gp80) and a
signal transducing subunit (gp130). IL-6 is able to bind gp80, but
does not bind to gp130 unless in the presence of gp80.
The cDNA for human gp80 has been isolated (Yamasaki et al., 1988,
Science 241), and was found to be 1407 bp in length. Human gp80
cDNA encodes a 468 amino acid protein, having a 19 amino acid
signal peptide and a domain of approximately 90 amino acids that is
similar to a domain in the immunoglobulin superfamily. The
cytoplasmic domain of approximately 82 amino acids lacks a
tyrosine/kinase domain, unlike other growth factor receptors. The
mature human protein has a calculated molecular weight of 51.6 kDa.
A soluble form of gp80 has been reported (Novick et al., 1989, J.
Exp. Med. 170) which arises from proteolytic cleavage of
membrane-bound gp80. This soluble receptor has been shown to bind
to IL-6 in solution (Yasukawa et al., 1990, J. Biochem. 108)
Extensive safety testing is required for an IL-6 or gp80 (IL-6R)
human therapeutic to be brought to the marketplace. Such safety
testing involves both in vivo safety testing in animal models as
well as the in vitro testing of these therapeutics. For example,
antibody based IL-6 and gp80 therapeutics may require the
generation of surrogate antibodies against an IL-6 or gp80 peptide
chain expressed by a particular model animal as well as significant
in vitro characterization of such surrogate antibodies. Such
surrogate generation and in vitro characterization may require the
use of IL-6 and gp80 polynucleotides and peptide chains from a
suitable model animal. Importantly, the identification of suitable
animal models for such safety testing requires the identification
of animal species capable of expressing a gp80 with high identity
and homology to human gp80 (SEQ ID NO:11).
Thus, a need exists for the identification of polynucleotides
encoding gp80s and gp80 peptide chains capable of being expressed
in an animal model suitable for the safety testing of IL-6 and gp80
therapeutics. A need also exists for related methods such as
methods of expressing peptide chains and testing the safety of an
IL-6 or gp80 therapeutic in an animal model identified as suitable
for safety assessment of IL-6 or gp80 therapeutics.
BRIEF DESCRIPTION OF THE DRAWINGS
FIG. 1 is a depiction of a gel showing RT-PCR products separated by
agarose gel electrophoresis and visualized under UV light,
including a band for cyno gp80.
FIG. 2 is a vector map of the polynucleotide (SEQ ID NO:1) encoding
the full-length mature cyno gp80 peptide chain.
FIG. 3 is a vector map of the polynucleotide (SEQ ID NO:3) encoding
the his-tagged full-length mature cyno gp80 peptide chain.
FIG. 4 is a vector map of the polynucleotide (SEQ ID NO:5) encoding
the extracellular domain of the mature cyno gp80 peptide chain.
FIG. 5 is a vector map of the polynucleotide (SEQ ID NO:7) encoding
the his-tagged extracellular domain of the mature cyno gp80 peptide
chain.
SUMMARY OF THE INVENTION
One aspect of the invention is an isolated polynucleotide
comprising a polynucleotide encoding cyno gp80 having the sequence
shown in SEQ ID NO:1 or a complementary sequence thereof. Another
aspect of the invention is an isolated polynucleotide comprising a
polynucleotide encoding the extracellular domain of cyno gp80
having the sequence shown in SEQ ID NO:5 or a complementary
sequence thereof.
Another aspect of the invention is a vector comprising an isolated
polynucleotide having the sequence shown in SEQ ID NOS:1 or 5.
Another aspect of the invention is a polypeptide comprising a
peptide chain having the mature, full-length cyno gp80 sequence set
forth in SEQ ID NO:2. Yet another aspect of the invention is a
polypeptide comprising a peptide chain having his-tagged, mature,
full-length cyno gp80 sequence set forth in SEQ ID NO:4. A further
aspect of the invention is a polypeptide comprising a peptide chain
having the extracellular domain of mature cyno gp80 sequence set
forth in SEQ ID NO:6. An additional aspect of the invention is a
polypeptide comprising a peptide chain having the his-tagged
extracellular domain of mature cyno gp80 sequence set forth in SEQ
ID NO:8. Also included in the invention is a polypeptide comprising
a peptide chain having the mature cyno gp80 sequence set forth in
SEQ ID NO:10 without the signal sequence.
Another aspect of the invention is a method for expressing a
peptide chain comprising the steps of providing a DNA or RNA
sequence coding for a polypeptide comprising the sequence set forth
in SEQ ID NOS:2, 6, or 10; providing the components of a cell free
expression system; initiating cell free expression from the RNA
provided; recovering the peptide chain; and confirming expression
of at least one peptide chain comprising the sequence set forth in
SEQ ID NOS:2, 6, or 10.
Another aspect of the invention is a method for determining if a
gp80 therapeutic causes adverse events comprising providing a gp80
therapeutic, a first cynomolgus monkey, and a second cynomolgus
monkey; administering the gp80 therapeutic to the first cynomolgus
monkey; and determining whether the first cynomolgus monkey is
presenting a deleterious symptom relative to the second monkey,
where presentation of a deleterious symptom by the first cynomolgus
monkey shows the gp80 therapeutic is unsafe and a lack of
presentation of a deleterious symptom by the first cynomolgus
monkey shows the gp80 therapeutic is safe.
DETAILED DESCRIPTION OF THE INVENTION
All publications, including but not limited to patents and patent
applications, cited in this specification are herein incorporated
by reference as though fully set forth.
As used herein and in the claims, the singular forms "a," "and,"
and "the" include plural reference unless the context clearly
dictates otherwise. Thus, for example, reference to "a peptide
chain" is a reference to one or more peptide chains and includes
equivalents thereof known to those skilled in the art.
Unless defined otherwise, all technical and scientific terms used
herein have the same meaning as commonly understood by one of
ordinary skill in the art to which an invention belongs. Although
any compositions and methods similar or equivalent to those
described herein can be used in the practice or testing of the
invention, exemplary compositions and methods are described
herein.
The present invention provides isolated cynomolgus monkey (Macaca
fascicularis) ("cyno") gp80 polynucleotides, vectors comprising
these polynucleotides, isolated host cells, peptide chains
obtainable from expression of these polynucleotides, methods for
expressing the peptide chains of the invention, and uses of these
are disclosed.
Importantly, the full-length cyno gp80 peptide chain sequence (SEQ
ID NO:2) is 97% identical, and 96.8% similar to the human gp80
(IL-6R) peptide chain (SEQ ID NO:11).
The compositions and methods of the invention can be used for a
variety of specific applications. The polynucleotides and vectors
of the invention are useful because they encode cynomolgus monkey
gp80 peptide chains and can be used to express these peptide
chains. These cyno gp80 peptide chains are, in turn, useful because
they can be used to increase or control inflammatory responses
after exposure to dsRNA or other gp80 ligands when they are
recombinantly over expressed or introduced by other means into a
host animal or tissue.
Peptide chains comprising the extracellular domain of cyno gp80 can
also be used as ligand sink type antagonists that bind available
gp80 ligands or gp80 associated proteins necessary for gp80
activation and thus control gp80 or IL-6-related activity. Cyno
gp80 peptide chains can also be used to generate therapeutic
antibodies for the positive or negative modulation of the activity
of cyno gp80 or gp80s from other sources. This is desirable because
agonist therapeutic antibodies can be used to increase activation
of cyno gp80 or other gp80s to help control inflammatory responses
while antagonist therapeutic antibodies can be used to decrease
activation of cyno gp80 or other gp80s to help control conditions
associated with gp80 receptor activation mediated inflammatory
responses. Cyno gp80 peptide chains can also be used in in vitro or
in vivo assays to identify other therapeutics, such as small
molecules and non-antibody biological therapeutics (e.g., catalytic
proteins) capable of modulating the activity of cyno gp80 or other
gp80s. The methods of expression disclosed are useful because these
methods permit the expression of cyno gp80 peptides. Other methods
disclosed are useful because they permit a safety assessment of a
gp80 therapeutic.
The term "polynucleotide" means a molecule comprising a chain of
nucleobases covalently linked by a sugar-phosphate backbone or
other equivalent covalent chemistry. Double and single stranded
DNAs and RNAs are typical examples of polynucleotides.
The term "complementary sequence" means a second isolated
polynucleotide sequence that is antiparallel to a first isolated
polynucleotide sequence and that comprises nucleobases
complementary to the nucleobases in the first polynucleotide
sequence. Typically, such "complementary sequences" are capable of
forming a double stranded polynucleotide molecule such as double
stranded DNA or double stranded RNA when combined under appropriate
conditions with the first isolated polynucleotide sequence.
The term "vector" means a polynucleotide capable of being
duplicated within a biological system or that can be moved between
such systems. Vector polynucleotides typically contain elements,
such as origins of replication or selection markers, that function
to facilitate the duplication or maintenance of these
polynucleotides in a biological system. Examples of such biological
systems may include cell, virus, animal, plant, and reconstituted
biological systems utilizing biological components capable of
duplicating a vector. The polynucleotides comprising a vector may
be DNA or RNA molecules or hybrids of these.
The term "expression vector" means a vector that can be utilized in
a biological system or a reconstituted biological system to direct
the translation of a peptide chain encoded by a polynucleotide
sequence present in the expression vector.
The term "peptide chain" means a molecule that comprises at least
two amino acid residues linked by a peptide bond to form a chain.
Large peptide chains of more than 50 amino acids may be referred to
as "polypeptides" or "proteins." Small peptide chains of less than
50 amino acids may be referred to as "peptides."
The term "antibody" means immunoglobulin or antibody molecules
comprising polyclonal antibodies, monoclonal antibodies including
murine, human, humanized and chimeric monoclonal antibodies and
antibody fragments, portions, or variants. Antibodies are secreted
proteins constitutively expressed and secreted by plasma cells.
Antibodies may be of any isotype such as IgG, IgA, or IgM and may
comprise antibody fragments such as Fab' fragments. An antibody may
also be a bispecific antibody that specifically binds two different
peptide chain epitopes.
Antibodies can be produced using plasma cells immortalized by
standard methods such as hybridoma generation or by transfection of
antibody heavy and/or light chain genes into an immortalized B
cell, such as a myeloma cell or other cell types, such as Chinese
hamster ovary (CHO) cells, plant cells and insect cells.
The term "monoclonal antibody" (mAb) as used herein means an
antibody (or antibody fragment, such as a Fab, single domain
antibody, etc.) obtained from a population of substantially
homogeneous antibodies. Monoclonal antibodies are highly specific,
typically being directed against a single antigenic determinant.
The modifier "monoclonal" indicates the substantially homogeneous
character of the antibody and does not require production of the
antibody by any particular method. For example, murine mAbs can be
made by the hybridoma method of Kohler et al., 256 Nature 495
(1975). Chimeric mAbs containing a light chain and heavy chain
variable region derived from a donor antibody (typically murine) in
association with light and heavy chain constant regions derived
from an acceptor antibody (typically another mammalian species such
as human) can be prepared by the method disclosed in U.S. Pat. No.
5,807,715. Humanized mAbs having CDRs derived from a non-human
donor immunoglobulin (typically murine) and the remaining
immunoglobulin-derived parts of the molecule being derived from one
or more human immunoglobulins, optionally having altered framework
support residues to preserve binding affinity, can be obtained by
the techniques disclosed in Queen et al., 86 Proc. Natl. Acad. Sci.
(USA) 10029 (1989) and Hodgson et al., 9 Bio/Technology 421
(1991).
Exemplary human framework sequences useful for humanization are
disclosed at, e.g., www.ncbi.nlm.nih.gov/entrez/query.fcgi;
www.ncbi.nih.gov/igblast; www.atcc.org/phage/hdb.html;
www.mrc-cpe.cam.ac.uk/ALIGNMENTS.php;
www.kabatdatabase.com/top.html; ftp.ncbi.nih.gov/repository/kabat;
www.sciquest.com; www.abcam.com;
www.antibodyresource.com/onlinecomp.html;
www.public.iastate.edu/.about.pedro/research_tools.html;
www.whfreeman.com/immunology/CH05/kuby05.htm;
www.hhmi.org/grants/lectures/1996/vlab;
www.path.cam.ac.uk/.about.mrc7/mikeimages.html;
mcb.harvard.edu/BioLinks/Immunology.html; www.immunologylink.com;
pathbox.wustl.edu/.about.hcenter/index.html;
www.appliedbiosystems.com; www.nal.usda.gov/awic/pubs/antibody;
www.m.ehime-u.ac.jp/.about.yasuhito/Elisa.html; www.biodesign.com;
www.cancerresearchuk.org; www. biotech.ufl.edu; www.isac-net.org;
baserv.uci.kun.nl/.about.jraats/links1.html;
www.recab.uni-hd.de/immuno.bme.nwu.edu; www.mrc-cpe.cam.ac.uk; www.
ibt.unam.mx/vir/V_mice.html; http://www.bioinf.org.uk/abs;
antibody.bath.ac.uk; www.unizh.ch;
www.cryst.bbk.ac.uk/.about.ubcg07s;
www.nimr.mrc.ac.uk/CC/ccaewg/ccaewg.html;
www.path.cam.ac.uk/.about.mrc7/humanisation/TAHHP.html;
www.ibt.unam.mx/vir/structure/stat_aim.html;
www.biosci.missouri.edu/smithgp/index.html; www.jerini.de; and
Kabat et al., Sequences of Proteins of Immunological Interest, U.S.
Dept. Health (1987), each entirely incorporated herein by
reference.
Fully human mAbs lacking any non-human sequences can be prepared
from human immunoglobulin transgenic mice by techniques referenced
in, e.g., Lonberg et al., 368 Nature 856 (1994); Fishwild et al.,
14 Nature Biotech. 845 (1996) and Mendez et al., 15 Nature Genetics
146 (1997). Human mAbs can also be prepared and optimized from
phage display libraries by techniques referenced in, e.g., Knappik
et al., 296 J. Mol. Biol. 57 (2000) and Krebs et al., 254 J.
Immunol. Meth. 67 (2001).
An antibody molecule or preparation "specifically binds" a given
antigen when it binds this antigen with higher affinity and in a
specific, as opposed to non-specific fashion, relative to a second
non-identical antigen. Stated differently, the "specific binding"
of an antibody molecule or preparation can be used to distinguish
between two different peptide chains.
A "fragment" is a peptide chain having an amino acid sequence that
comprises a portion, but not all, of any amino acid sequence of any
peptide chain of the invention. Fragments can include, for example,
truncated peptide chain having a portion of an amino acid sequence
corresponding to a signal peptide, extracellular domain,
transmembrane domain, or cytoplasmic domain, or variants thereof,
such as a continuous series of residues that includes a
heterologous amino- and/or carboxy-terminal amino acid sequence.
Degradation forms of the peptide chains of the invention produced
by, or in, a host cell are also included. Other exemplary fragments
are characterized by structural or functional attributes such as
fragments that comprise alpha-helix or alpha-helix forming regions,
beta-sheet or beta-sheet forming regions, turn or turn-forming
regions, coil or coil-forming regions, hydrophilic regions,
hydrophobic regions, alpha-amphipathic regions, beta-amphipathic
regions, flexible regions, surface-forming regions, substrate
binding regions, extracellular regions and high antigenic index
regions. Importantly, the peptide chains of the invention can be
used or provided as fragments.
A "variant peptide chain" is a second peptide chain in which amino
acid substitutions, insertions, deletions or combinations thereof
have been made relative to a first peptide chain. Naturally
occurring, modified or atypical amino acids can be used for
substitutions and insertions.
A "variant polynucleotide" is a second polynucleotide in which
nucleic acid residue substitutions, insertions, deletions, or
combinations thereof have been made relative to a first
polynucleotide sequence. Naturally occurring or modified
nucleobases can be used for substitutions and deletions. The
various polynucleotides encoding the amino acid sequence set forth
in SEQ ID NOS:2, 4, 6, 8 and 10 are additional exemplary variant
polynucleotides relative to the polynucleotide having the
nucleotide sequences set forth in SEQ ID NOS:1, 3, 5, and 7.
The term "gp80 therapeutic" means a molecule or preparation that is
believed to provide a therapeutic benefit and is believed to
provide that therapeutic benefit, in part, through the activity of
a gp80. Such gp80s may comprise the peptide chains of the invention
or be generated using the peptide chains of the invention. Examples
of gp80 therapeutics include cyno gp80 agonists, antibodies or
other antagonists to cyno gp80, known cyno gp80 ligands, such as
IL-6, that produce the therapeutic benefits of increased
anti-inflammatory activity and the like.
The term "deleterious symptom" means any symptom presented by an
animal that indicates harm to the animal has occurred.
One aspect of the invention is an isolated polynucleotide
comprising a polynucleotide having the sequence set forth in SEQ ID
NOS:1, 3, 5, or 7 or a complementary sequence thereof. The
polynucleotide sequences set forth in SEQ ID NOS: 1, 3, 5, and 7
encode peptide chain comprising the predicted mature form of cyno
gp80, the his-tagged mature form, extracellular domain of cyno gp80
and his-tagged extracellular domain.
The polynucleotides of the invention may be produced by chemical
synthesis, such as solid phase polynucleotide synthesis on an
automated polynucleotide synthesizer. Alternatively, the
polynucleotides of the invention may be produced by other
techniques, such as PCR based duplication, vector based
duplication, or restriction enzyme based DNA manipulation
techniques. Techniques for producing or obtaining polynucleotides
of a given known sequence are well known in the art.
The polynucleotides of the invention may also comprise at least one
non-coding sequence, such as transcribed but not translated
sequences, termination signals, ribosome binding sites, mRNA
stabilizing sequences, introns and polyadenylation signals. The
polynucleotide sequences may also comprise additional sequences
encoding additional amino acids. These additional polynucleotide
sequences may, for example, encode a marker or tag sequence such as
a hexa-histidine peptide, as described in Gentz et al., 86 Proc.
Natl. Acad. Sci. (USA) 821 (1989) or the HA peptide tag as
described in Wilson et al., 37 Cell 767 (1984) which facilitate the
purification of fused polypeptides.
The present invention also includes vectors comprising an isolated
polynucleotide having the sequence set forth in SEQ ID NOS:1, 3, 5,
and 7. Expression vector maps including SEQ ID NO:1, encoding SEQ
ID NO:2, and including SEQ ID NO:3, encoding SEQ ID NO:4, the
his-tagged cyno gp80, are shown in FIGS. 2 and 3, respectively. The
vector shown in FIG. 2 is a polynucleotide (DNA) expression vector
designated pDR1981 that encodes a peptide chain comprising
full-length cyno gp80 (SEQ ID NO:2). The vector shown in FIG. 3 is
a polynucleotide (DNA) expression vector designated
pUNDER/cynoIL6R.5 that encodes a peptide chain comprising the
his-tagged full-length cyno gp80 (SEQ ID NO:2). The His tag is
6.times. with G-S residues inserted between the gp80 sequence and
tag during cloning.
Expression vector maps including SEQ ID NO:5, encoding SEQ ID NO:6,
the extracellular domain of cyno gp80, and including SEQ ID NO:7,
encoding SEQ ID NO:8, the his-tagged extracellular domain of cyno
gp80, are shown in FIGS. 4 and 5, respectively. The vector shown in
FIG. 4 is a polynucleotide (DNA) expression vector designated
pDR1983. The vector shown in FIG. 5 is a polynucleotide (DNA)
expression vector designated pUNDER/cynoIL6R ECD.5. The His tag is
6.times. with G-S residues inserted between the gp80 sequence and
tag during cloning.
The vectors of the invention are useful for maintaining
polynucleotides, duplicating polynucleotides, or driving expression
of a peptide chain encoded by a vector of the invention in a
biological systems--including reconstituted biological systems.
Vectors may be chromosomal-, episomal- and virus-derived such as
vectors derived from bacterial plasmids, bacteriophages,
transposons, yeast episomes, insertion elements, yeast chromosomal
elements, baculoviruses, papova viruses such as SV40, vaccinia
viruses, adenoviruses, fowl pox viruses, pseudorabies viruses,
picronaviruses and retroviruses and vectors derived from
combinations thereof, such as cosmids and phagemids.
The vectors of the invention can be formulated in microparticles,
with adjuvants, with lipid, buffer or other excipients as
appropriate for a particular application.
In one embodiment of the invention the vector is an expression
vector.
Expression vectors typically comprise nucleic acid sequence
elements that can control, regulate, cause or permit expression of
a peptide chain encoded by such a vector. Such elements may
comprise transcriptional enhancer binding sites, RNA polymerase
initiation sites, ribosome binding sites, and other sites that
facilitate the expression of encoded peptide chains in a given
expression system. Such expression systems may be cell based, or
cell free systems well known in the art. Nucleic acid sequence
elements and parent vector sequences suitable for use in the
expression of encoded peptide chains are also well known in the
art.
Another embodiment of the invention is an isolated host cell
comprising a vector of the invention.
An exemplary plasmid derived expression vector useful for
expression of the polypeptides of the invention comprises an E.
coli origin of replication, an aph(3')-1a kanamycin resistance
gene, HCMV immediate early promoter with intron A, a synthetic
polyA sequence and a bovine growth hormone terminator. Another
exemplary plasmid derived expression vector comprises an E. coli
origin of replication, an ant(4')-1a kanamycin resistance gene,
Rous sarcoma virus long terminal repeat sequences, HCMV immediate
early promoter and an SV40 late polyA sequence.
Representative host cell examples include Archaea cells; bacterial
cells such as Streptococci, Staphylococci, Enterococci, E. coli,
Streptomyces, cyanobacteria, B. subtilis and S. aureus; fungal
cells such as Kluveromyces, Saccharomyces, Basidomycete, Candida
albicans or Aspergillus; insect cells such as Drosophila S2 and
Spodoptera Sf9; animal cells such as CHO, COS, HeLa, C127, 3T3,
BHK, 293, CV-1, Bowes melanoma and myeloma; and plant cells, such
as gymnosperm or angiosperm cells. The host cells in the methods of
the invention may be provided as individual cells, or populations
of cells. Populations of cells may comprise an isolated or cultured
population of cells or cells present in a matrix such as a
tissue.
Introduction of a polynucleotide, such as a vector, into a host
cell can be effected by methods well known to those skilled in the
art from laboratory manuals such as Davis et al., Basic Methods in
Molecular Biology, 2.sup.nded., Appleton & Lange, Norwalk,
Conn. (1994) and Sambrook et al., Molecular Cloning: A Laboratory
Manual, 3.sup.ded., Cold Spring Harbor Laboratory Press, Cold
Spring Harbor, N.Y. (2001). These methods include calcium phosphate
transfection, DEAE-Dextran mediated transfection, microinjection,
cationic lipid-mediated transfection, electroporation,
transduction, scrape loading, ballistic introduction and
infection.
The present invention comprises an isolated peptide chain having
the sequence set forth in SEQ ID NO:2. SEQ ID NO:2 is a peptide
chain comprising the predicted mature form of cyno gp80. The
present invention also comprises an isolated peptide chain
comprising a peptide chain having the sequence set forth in SEQ ID
NO:4. SEQ ID NO:4 is a peptide chain comprising the His-tagged
predicted mature form of cyno gp80.
The present invention comprises an isolated peptide chain having
the sequence set forth in SEQ ID NO:6. SEQ ID NO:6 is a peptide
chain comprising the predicted extracellular domain of the mature
form of cyno gp80. The present invention also comprises an isolated
peptide chain comprising a peptide chain having the sequence set
forth in SEQ ID NO:8. SEQ ID NO:8 is a peptide chain comprising the
His-tagged extracellular domain of the predicted mature form of
cyno gp80. The present invention further comprises an isolated
peptide chain comprising a peptide chain having the sequence set
forth in SEQ ID NO:10. SEQ ID NO:10 is a peptide chain comprising
the predicted mature form of cyno gp80 without the 19-amino acid
signal sequence.
The peptide chains of the invention may be produced by chemical
synthesis, such as solid phase peptide syntheses, on an automated
peptide synthesizer. Alternatively, the peptide chains of the
invention can be obtained from polynucleotides encoding these
peptide chains by the use of cell free expression systems such as
reticulocyte lystate based expression systems, wheat germ extract
based expression systems, and Escherichia coli extract based
expression systems. The peptide chains of the invention can also be
obtained by expression and isolation from cells harboring a nucleic
acid sequence of the invention by techniques well known in the art,
such as recombinant expression of easily isolated affinity labeled
peptide chains. Those skilled in the art will recognize other
techniques for obtaining the peptide chains of the invention.
The peptide chains of the invention may comprise fusion peptide
chains comprising a peptide chain of the invention fused with
second peptide chain. Such second peptide chains may be leader or
secretory signal sequences, a pre- or pro- or prepro-protein
sequence, as well as naturally occurring, or partially synthetic
sequences derived in part from a naturally occurring sequence or an
entirely synthetic sequence. Secretory signal or leader peptide
chain sequences may be selected to direct secretion of the peptide
chains of the invention into the lumen of the endoplasmic reticulum
or extracellular environment; such peptide chain sequences may be
heterologous or endogenous to any peptide chain from a cynomolgus
monkey or comprise hybrids of these.
The peptide chains of the invention can also be formulated in a
pharmaceutically acceptable carrier or diluent. A variety of
aqueous carriers may be employed, e.g., 0.4% saline, 0.3% glycine
and the like. These solutions are sterile and generally free of
particulate matter. These solutions may be sterilized by
conventional, well-known sterilization techniques (e.g.,
filtration). The compositions may contain pharmaceutically
acceptable auxiliary substances as required to approximate
physiological conditions, such as pH adjusting and buffering
agents. The concentration of the peptide chains of the invention in
such pharmaceutical formulation can vary widely, i.e., from less
than about 0.5%, usually at or at least about 1% to as much as 15
or 20% by weight and will be selected primarily based on fluid
volumes, viscosities and other factors, according to the particular
mode of administration selected.
The peptide chains and nucleic acids of the invention can also be
provided in the form of a pharmaceutical preparation, such as a
vaccine for eliciting an immune response, that can be provided in
unit dose forms. The appropriate therapeutically effective dose can
be determined readily by those of skill in the art. A determined
dose may, if necessary, be repeated at appropriate time intervals
selected as appropriate by a physician or other person skilled in
the relevant art (e.g., nurse, veterinarian, or veterinary
technician) during the treatment period.
The peptide chains of the invention can be lyophilized for storage
and reconstituted in a suitable carrier prior to use. This
technique has been shown to be effective with conventional protein
preparations. Lyophilization and reconstitution techniques are well
known in the art.
Another embodiment of the invention is a method for expressing a
peptide chain comprising the steps of providing a host cell of the
invention; culturing the host cell under conditions sufficient for
the expression of at least one peptide chain comprising the
sequence set forth in SEQ ID NOS:2, 4, 6, 8, or 10; recovering the
peptide chain; and, optionally, confirming expression of at least
one peptide chain comprising the sequence set forth in SEQ ID
NOS:2, 4, 6, 8, or 10.
Host cells can be cultured under any conditions suitable for
maintaining or propagating a given type of host cell and sufficient
for expressing a peptide chain. Culture conditions, media, and
related methods sufficient for the expression of peptide chains are
well known in the art. For example, many mammalian cell types can
be aerobically cultured at 37.degree. C. using appropriately
buffered DMEM media while bacterial, yeast and other cell types may
be cultured at 37.degree. C. under appropriate atmospheric
conditions in LB media.
In the methods of the invention the expression of a peptide chain
can be confirmed using a variety of different techniques well known
in the art. For example, expression of a peptide chain can be
confirmed using detection reagents, such as antibodies or receptor
ligands, specific for an expressed peptide chain. Antibodies that
specifically bind to the cyno gp80 peptide chains of the invention
are one example of such reagents. gp80 receptor ligands, such as
IL-6, dsRNA or poly(I:C) that bind gp80, are additional examples of
such reagents. Detection reagents may be detectably labeled by
conjugation or incorporation of a radiolabel, fluorophore,
chromophore or other detectable molecule to, or into, the detection
reagent.
Alternatively, the expression of a cyno gp80 peptide chain of the
invention can be confirmed by assaying for a biological activity
associated with activation of gp80s, such as an inflammatory
response. Such assays may also utilize reporter gene constructs
responsive to gp80 activation.
Peptide chain expression can also be confirmed by identification of
a peptide chain with the physical characteristics of a peptide
chain of the invention in a preparation of peptide chains. For
example, SDS-PAGE techniques and other well-known protein
characterization techniques utilizing criteria such as, for
example, protein molecular weight or isoelectric point can be used
to confirm expression of the peptide chains of the invention.
Protein purification techniques such as ammonium sulfate or ethanol
precipitation, acid extraction, high-performance liquid
chromatography, anion or cation exchange chromatography,
phosphocellulose chromatography, hydrophobic interaction
chromatography, affinity chromatography, hydroxyapatite
chromatography and lectin chromatography can also be used to
confirm expression of a peptide chain of the invention.
Importantly, in the methods of the invention the peptide chain
expressed need not be isolated. Consequently, expression of a
peptide chain may be confirmed to have occurred on, or in, a cell,
or in a mixture of peptide chains for example. Flow cytometry based
techniques such as fluorescence activated cell sorting (FACS) may
also be used, when appropriate, to confirm expression of a peptide
chain by a cell. As discussed above peptide chain expression may be
confirmed using any suitable technique known in the art.
Another embodiment of the invention is a method for expressing a
peptide chain comprising the steps of providing a polynucleotide of
the invention capable of being transcribed into an RNA coding for
at least one peptide chain comprising the sequence set forth in SEQ
ID NOS:2, 4, 6, 8, or 10; providing the components of a cell free
expression system; initiating cell free expression from the
polynucleotide provided; recovering the peptide chain; and,
optionally, confirming expression of at least one peptide chain
comprising the sequence set forth in SEQ ID NOS:2, 4, 6, 8, or
10.
Techniques for transcribing a polynucleotide into an RNA, obtaining
an RNA coding for a peptide chain, or initiating cell free
expression are well known in the art and reagent kits for
accomplishing these steps are commercially available from a variety
of sources.
In another embodiment of the method of the invention the cell free
expression system is selected from the group consisting of a
reticulocyte lystate based expression system, a wheat germ extract
based expression system, and an Escherichia coli extract based
expression system.
Another embodiment of the invention is a method for expressing a
peptide chain comprising the steps of providing an RNA coding for
at least one peptide chain comprising the sequence set forth in SEQ
ID NOS:2, 4, 6, 8, or 10; providing the components of a cell free
expression system; initiating cell free expression from the RNA
provided; recovering the peptide chain; and, optionally, confirming
expression of at least one peptide chain comprising the sequence
set forth in SEQ ID NOS:2, 4, 6, 8, or 10.
In another embodiment of the method of the invention the cell free
expression system is selected from the group consisting of a
reticulocyte lystate based expression system, a wheat germ extract
based expression system, and an Escherichia coli extract based
expression system.
Another embodiment of the invention is a peptide chain produced by
the methods of invention. Such peptide chains may comprise
post-translational modifications including glycosylation or
phosphorylation for example. Such peptide chains may also comprise
alternative peptide chain forms such as splice variants, truncated
forms, or proteolytically modified forms.
Another embodiment of the invention is an antibody that
specifically binds a peptide chain of the invention. The peptide
chains of the invention can be used to produce polyclonal or
monoclonal antibodies against cyno gp80. Techniques for making
murine, chimeric, humanized and fully human monoclonal antibodies
using protein or nucleic acid immunization are routine and well
known to those skilled in the art. Additional discussion and
description of such techniques can be found above.
Another embodiment of the invention is a monoclonal antibody that
specifically binds a peptide chain of the invention.
Another aspect of the invention is a method for determining if a
gp80 therapeutic is safe or unsafe comprising providing a gp80
therapeutic, a first cynomolgus monkey, and a second cynomolgus
monkey; administering the gp80 therapeutic to the first cynomolgus
monkey; and determining whether the first cynomolgus monkey is
presenting a deleterious symptom relative to the second monkey,
where presentation of a deleterious symptom by the first cynomolgus
monkey shows the gp80 therapeutic is unsafe and a lack of
presentation of a deleterious symptom by the first cynomolgus
monkey shows the gp80 therapeutic is safe.
In the methods of the invention the first and second cynomolgus
monkey provided should be equivalent with regard to the
presentation of deleterious symptoms. Stated differently both
animals should be presenting either no deleterious symptoms or the
same deleterious symptoms when they are provided.
In the methods of the invention gp80 therapeutics can be
administered by any route appropriate, such as parenterally,
subcutaneously, intravenously, etc. Examples of gp80 therapeutics
suitable for use in the method of the invention include, for
example, known gp80 ligands, such as IL-6 peptides, dsRNA or
poly(I:C) and small molecule and biological therapeutics that
impact binding of IL-6 to gp80 and resulting signalling.
In the methods of the invention the determination of whether the
first cynomolgous monkey is presenting a deleterious symptom
relative to the second cynomolgous monkey is readily accomplished.
For example, a person of ordinary skill in the art such as a
veterinarian, veterinarian's assistant, animal technician, or
research scientist can determine if a symptom presented by an
animal is deleterious. Examples of deleterious symptoms include
death, coma, seizures, fever, organ failure, tissue abnormalities,
impaired organ function, impaired tissue function, cancers, tumors,
ulcers, bleeding, infections and the like.
In one embodiment of the method of the invention the gp80
therapeutic is an antibody.
The present invention will now be described with reference to the
following specific, non-limiting examples.
EXAMPLE 1
Isolation, Cloning and Sequencing of Polynucleotides Encoding
Full-Length Cynomolgus gp80
Kidney tissue from a Macaca fascicularis (cyno) monkey was obtained
from BioChain (Hayward, Calif.). RNA was isolated from kidney
tissue from Macaca fascicularis (cyno) and reverse transcribed
using the Superscript III kit (Invitrogen) into a cDNA pool.
The cynomolgus gp80 gene was then amplified from this cDNA by
RT-PCR and sequenced. Using the predicted nucleotide sequence for
rhesus gp80 (Genbank Accession XM.sub.--001114404), oligos were
designed to 5' (nucleotides 383-403 (underlined)) and 3'
(nucleotides 1850-1871 (underlined)) untranslated regions (Table 1,
SEQ ID NO:9 in which the coding region is from nucleotides
423-1829). It was assumed that a high degree of similarity exists
between rhesus and cyno monkeys such that the primers would anneal
to the cyno sequence. Using these oligos, RT-PCR was performed
using the cyno cDNA pool as a template for amplification. A
fragment of approximately 1.5 kb was isolated and subcloned using
the TOPO-TA kit (Invitrogen) (FIG. 1). Plasmid DNAs from 8
transformants were isolated and sequenced. The confirmed sequence
of cyno gp80 is shown in Table 2.
The sequence of cynomolgus gp80 protein is 97% identical to the
human gp80 amino acid sequence. A sequence comparison of the human
and cyno gp80 nucleotide sequences is shown in Table 3, and amino
acid sequences in Table 4. This high degree of conservation
suggests that the cynomolgus monkey is a relevant toxicology study
animal for the in vivo evaluation of compounds that target gp80 (or
alternatively IL-6).
RT-PCR was performed using oligos based on the rhesus 5' and 3' UTR
on a cDNA pool derived from cyno kidney tissue as a template for
amplification. PCR products were separated by agarose gel
electrophoresis and visualized under UV light. This is shown in
FIG. 1. A fragment of approximately 1.5 kb was expected; the band
indicated by the red arrow was isolated (NTC=No Template Control;
MW=Molecular Weight marker).
It will be clear to one of ordinary skill in the art that the
invention now being fully described can be practiced otherwise than
as particularly described in the foregoing description and
examples. Numerous modifications and variations of the present
invention are possible in light of the above teachings and,
therefore, are within the scope of the appended claims and
equivalents.
Tables
TABLE-US-00001 TABLE 1 Nucleotide Sequence of rhesus gp80 (SEQ ID
NO: 9) 1 GGCGGTCCCCTGTTC TCCCCGCTCAGGTGC GGCGCTGTGGCAGGA
AGCCACCCCCTCGGT CGGCCGGTGCGCGGG 76 GCTGTTGCGCCATCC GCTCCAGCTTTCGTA
ACCGCACCCTGGGAC GGCCCAGAGACGCTC CAGCGCGAGTTCCTC 151 AAATGTTTTCCTGTG
TTGCCAGGACCGTCC GCCGCTCTGAGTCAT GTGCGAGTGGGAAGT CTCACTGACACTGAG 226
ACCGGCCAGTGGGAG CGGAGCCGAGCGCGG CGCGGGGCTGAGGGA CTCGCAGTGTATATA
GAGCGCCGGGCTCCT 301 GCGATGGGGGCTGCC CCCGGAGACTGAGCC AGCCTGCCCGCCCAC
CGCCCCGCCCCTGCC GCCCGGTTCCCACCA 376 GCCTGTCCGCCTCTG CGGGACCATGGAGCG
GTAGCCGAGGAGGAA GCATGCTGGCCGTCG GCTGCGCGCTGCTGG 451 CTGCCCTGCTGGCCG
CGCCGGGGGCGGCGC TGGCCCCGGGGGGCT GCCCTGCGCAGGAGG TGGCGAGAGGTGTGC 526
TGACCAGTCTGCCAG GAGACAGCGTGACTC TGACCTGCCCAGGGG GAGAGCCGGAAGACA
ATGCCACTGTTCACT 601 GGGTTCTCAGGAAGC CAGCTGTAGGCTCCC ACCTCAGCAGATGGG
CTGGCGTGGGAAGGA GGCTGCTGCTGAGGT 676 CGGTGCAGCTCCATG ACTCTGGAAACTATT
CATGCTACCGGGCCG GCCGCCCGGCTGGAA CTGTGCACTTGCTGG 751 TGGATGTTCCCCCCG
AGGAGCCCCAGCTCT CCTGCTTCCGGAAGA GCCCCCTCAGCAACG TTGTTTGTGAGTGGG 826
GTCCTCGGAGCACCC CATCTCCGACGACCA AGGCTGTGCTGTTGG TGAGGAAGTTTCAGA
ACAGTCCGGCCGAAG 901 ACTTCCAGGAGCCGT GCCAGTATTCCCAGG AGTCCCAGAAGTTCT
CCTGCCAGTTGGCAG TCCCGGAGGGAGACA 976 GCTCTTTCTACATAG TGTCCATGTGCGTCG
CCAGTAGTGTCGGGA GCAAGCTCAGCAAAA CTCAGACCTTTCAGG 1051
GTTGTGGAATCTTGC AGCCTGATCCGCCTG CCAACATCACAGTCA CTGCCGTGGCCAGAA
ACCCCCGCTGGCTCA 1126 GTGTCACCTGGCAAG ACCCCCACTCCTGGA
ACTCATCTTTCTACA GACTACGGTTTGAGC TCAGATATCGAGCTG 1201
AACGGTCAAAGACAT TCACAACATGGATGG TCAAGGACCTCCAGC ATCACTGTGTCATCC
ACGACGCCTGGAGCG 1276 GCCTGAGGCACGTGG TGCAGCTTCGTGCCC
AGGAGGAGTTCGGGC AAGGCGAGTGGAGCG AGTGGAGCCCGGAGG 1351
CCATGGGCACGCCTT GGACAGAATCCAGGA GTCCTCCAGCTGAGA ACGAGGTGTCCACCC
CCACGCAGGCACCTA 1426 CTACTAATAAAGATG ATGATAATATTCTCT
CCAGAGATTCTGCAA ATGCGACAAGCCTCC CAGTGCAAGATTCTT 1501
CTTCGGTACCACTGC CCACATTCCTGGTTG CTGGAGGGAGCCTGG CGTTCGGAACGCTCC
TGTGCATTGCCATTG 1576 TTCTGAGGTTCAAGA AGACGTGGAAGCTGC
GGGCTCTGAAGGAAG GCAAGACAAGCATGC ACCCGCCGTATTCTT 1651
TGGGGCAGCTGGTCC CAGAGAGGCCGCGAC CCACCCCAGTGCTTG TTCCTCTCATCTCCC
CACCAGTGTCCCCCA 1726 GCAGCCTGGGGTCTG ACAACACCTCGAGCC
ACAACCGACCAGATG CCAGGGACCCACGGA GCCCTTACGACATCA 1801
GCAATACAGACTACT TCTTCCCCAGATAGT TGGCCAGGGGGCACT AGCAGGCTGGACCCT
GTGGATGACAGAGCA 1876 CAAACGGGCTCAGCA AAGGATGCTTCTTAC
TGCCATGCCAGCTTA TCTCAGGGGAGTGGG GCCTTTGGCTTCACG 1951
GAAGAGCCTTGCGGA AGGTTCGACACCAGG GGAAAATCAGCCTGC TCCAGCTGTTCAGCT
GGTTGAGATTTCAAA 2026 CCTCCCTTTCCAAAT GTCTGGCTTAAAGGG
GTTAGAGTGAACTTG GGCCACTGTGAAGAG AAGCATGTCAAGACT 2101
CTTTGGACATTCAAC ACGGACACTCAAAAG CTGGGCAGCTTGGTG GGGGCCTCAGTGTGG
AGAAGCGGCTGGCAG 2176 CCCACCCCCCAACAC CTCTGCACAAGCTGC
GCCCTCAGGCAGGGG GGGCGGATTTCCAGC CAAAGCCTCCTTCAG 2251
CCGCCACGCTCCTGG CCCACTGCATCATTT CATCTTCCAGCTCAA ACTCCTAAAACCCAA
GTGCCTTTGCAAATT 2326 CTGTTTTTCTGAGCC TGGGGACGGCTTTTA
CTTAAACCGCCAAGG CTGGGGAAAGAAGCT CTCTCCTCCCTTTCT 2401
TCCCCACAGTTGAAA AACAGCTGAGGGTGG GTGGGTGAATAATAC AGTATCTCAGGGCCT
GGTCGTTTTCAACGG 2476 AATTATAATTAGTTC CTCATTAGCATTTTG
CTAAATGTGAATGAT AATCCTAGGCATTTG CTGAATACAGAGGCA 2551
ACTGCATTGGCTTTG GGTTGCAGGACCTCA GGTGAGAAGCAGAGG AAGGAGAGGAGAGGG
GCACAGGGTCTCCAC 2626 CATCCCCTGTAGAGT GGGAGCTGCGCGGGG
GATCACAGCCTCTGA AAACCAATGTTCTCT GTTCTCCACCTCCCA 2701
CAAAGGAGAGCTGGC AGCAGGGAGGGCTTC TGCCAGTGCTGAGAT CAAAACTGTTTTACT
GCAGCTTTGTTTGTT 2776 GTCAGCCGAACCTGG GTAACTAGGGAAGAT
AATATTAAGGAAGAC AATGTGAAAAGAAAA ATGAGCCCGGCAAGA 2851
ATGCATTTTAACTTG GTTTTTAAAAAACTG CTGACTGTTTTCTCT TGAGAGGGTGGAATA
TCCAATATGCGCTGT 2926 GTCAGCATAGAAGTA ACTTACTTAGGTGTG
GGGGAAGCACCATAA CTTTGTTTAGCCCAA AACTAAGTCAAGTGA 3001
AAAAGGAGGAAGAGA AATAATATTTTTCCT GCCAGGCATGGTGGT TCACGCCTGTAATCC
CAGCACTCTGGGAGG 3076 TCGAGGCGGGACGAT CACTTGAGTCCAGGA
GTTTGAGACCAGCCT GGGCAATGTGGTAAA ACCTCATCTCGACAA 3151
AAAGCATAAAAATTA GCCAGGTATGGTAGA GTGCACCTGAAGTCC CAGTTAGTTGGGAGG
CTGAGGTGGGAGGAT 3226 CTCTTGAGCCTGGGA GGTCAAGGCTGCAGT
GAGCCGAGATTGCAC CACTGCACTCCAGCC TGGGTGACTGAGCAA 3301
GTGAGACCCTGTCTC AAAAAAGAAAAGGAA AAAGAAAAGAAAAAA TATTTTCCCTGTTAG
AGAAGAGATTGTGGT 3376 TTCATTGTGTATTTT GTTTTTGTCTTAAAA
AGTGGAAAAATAGCC TGCCTCTTCTCTACT CTAGGGAAAAACCAG 3451
TGTGTGACTACTCCC CCAGGCGGTTATGGA GAGGGCGTCCGGTCC CTGTCCCAGTGCTGA
GAAGGGAGCCTCCCA 3526 CGACTACCCGGCAGG GTCCTAGAAATTCCC
CACCCTGAAAGCCCT GAGCCTTCTGCTATC AAAGGGGCAGGTAAA 3601
AATCCCATTTAAAAA AAATCCCTTACCTCG GTGCCTTCCTCTTTT TATTTAGCTCCTTGA
GTTGATTCAGCTCTG 3676 CAAGAATTGAAGCAG AACTAAATGTCTAAT
TGTAACACCGTGATT AACCACTTCAGCTGA CTTTTCTGCCCGAGC 3751
TTTGAAAATTCAGTG GTGTTAGTGGTTACC CAGTTAGCTCTCAAG TTATCAGGGTACTCC
ACAGCGGGGATATAC 3826 CAGACCACAAAACCT TTCTAATACTCTACC
CTCTTAGAAAAACAG CCACCATCACCAGAC AGGTGCAAAAGGAGG 3901
AAAGTGACCATGTTT TGTTTACCGTTTTCC AGGTTTAAGCTGTTA CTGTCTTCAGCAAGC
CGTGCTTTTCATTGC 3976 TGGGTTTGTCTGTAG ATTTTAGACCCTATT
GCTGCTTGAGGCACC TCATCTTAAGTTGGC AAAAAGGCAGGACGG 4051
CTGGGTGTGGTGGCT CACGCCTGTAATCCT AGCACTTTGGGAGGC CGAGGTGGGAGGATT
GCTTGAGCTCAGGAA 4126 TTTGAGACCAACCTG GGTAACATAGTGAGA
TACCATCTCTATTAT AAACAATAACATTTA AGGAAAAAAAAAGGC 4201
AGGCAGGTGGTTATG GTGGTTCCCTCCCAT CCTGCTGCATAAAGT TTCTGAGACTTGAGA
ACAGCAAAAATGCTG 4276 TTAAAGGGAAATATT AAGAATGAGAATCTG
CATGAAGGGTGATTA TGTGCCCACAGTTAA TTCTTTATACCGTTT 4351
TACCCACATGTGGTA TTACCGAAGCCGGGC AGAACCATGCTAGCG GAAGATATGAAATTC
AGATAGCTCATTATT 4426 GCCAAGAGCTAGGCA GCTTTGATCTCCAAA
TTGTTATTGCTTTCA TTTTTATTGTAATGG AACTGCTTTTTTTTT 4501
TTTTTTTTTTTTTTT TGCTTTTTTTTCTTT GTTTTGTTTTTGTAG TGAAGAGGGTTTTTT
TCCCTTTATTTTTCA 4576 TAAGCTACTGTAAAT GAAGAAAAAGTGTCT
TCTCTGGGCTGTAGG CCTGGCTCAGTGTAC ACAGGTATACATCCT 4651
AAGCTCTCTCTGTTC TCTAATTTGTGGTGA CTGAATATGTGTCGC AATCCACGGGGCATT
TCTACCTGTATTTCT 4726 GCAGCACCCCCACTG CCTTGAGTCCCCAGC
AGTGCTGTTATTTGC CTAATACCTGTAGCC ATCTGCCACACAGCC 4801
AGACATGAAACGCTG GGACAGAGACCATTT AGATTAAATACAACA GCTTATCTTGCTGGG
TGGGGAAAGTAAAAA 4876 ATATGCTGGTTCAAG GTCTAAAGTAAAATG
ATAAATAATGTTTGT AGCATTAATGAAATA TTTTCAAGAAATGTG 4951
TCCGGGGGTAGCATT GGCTATGCTGACGAG GCCTTTGGTAACTCA GAAAGCTCTTGGCCC
CGATGGCGACTTGCC 5026 CTTGCACTTTCTTTA TCAGGCTCTGAGCTC
ACACGGAGCCTCTGG CATTTCCCTGCTGTC TTGGGAGAAAGGAAA 5101
CTGGTTGTGGCGGCA GGGTGTGGAATCTGC TGCTGGAACCAGGCT GGAAGCCCACCTGGT
AGTGAACAGGGCCCA 5176 GCGGGGCAGGCTAGG AGTGTTGTGGTCTAT
GGGTTTGTGTCCTGG AGAATGTTCAAGAAT GTCTTCTTGGCTGCT 5251
TTGGTGCTGAGCTCT GTTATCTCACAGCAC GTCCTGAAGGCTAAC CCAGGTGGGGAGGAT
GCTGACACCAGCTCC 5326 AGGTGGAGTTGGTGA GAAATCTGTCTTAAC
TTGGAGATGCAGGGG CAACCTGTGACCCTT TGAGGCAAGAGCCCT 5401
GCACCCAGCTGTCCC GTGCAGCCGTGGGCA GGGGGCTGCACATGG AGGGGCAGGCGGGCC
AGTTCAGGGCCAGTT 5476 CAGTGCCCTGTAAGG GCCCTTCAGCCTCCT
GTCCTCTGTGCGGCT GGGCGCCAGCACCAG GGAGTTTCTATGGCA 5551
ACCTTAGTGATTATT AAGGAACATTGTCAG TTTTATGAACATATG CTCAAATGAAATTCT
ACTTTAGGAGGAAAG 5626 GATTGGAACAGCATG TTGCAAGGCTGTTAA
TTAACAGAGAGACCT TATTGGATGGAGATC ACATCTGTTAAATAG 5701
AATACCTCAACTCTA CGTTGTTTTCTTGGA GATAAATAATAGTTT CAAGTTTTTGTTTGT
TTGTTTTACCTAATT 5776 ACCTGAAAGCAAATA CCAAAGGCTGATGTC
TGTATATGGGGCAAA GGGTCAGTATATTTT TCAGTGTTTTTTTTT 5851
CTTTTACAAGCTATT TTGCATTTAAAGTGA ACATTGTAAATGTTT GTAATAAATGATTTT
TAAAAATACA
TABLE-US-00002 TABLE 2 cDNA and amino acid sequence of cyno gp80.
The encoded protein sequence (SEQ ID NO: 2) is shown above the
nucleotide sequence (SEQ ID NO: 1). M L A V G C A L L A A L L A T 1
ATGCTGGCC GTCGGCTGC GCGCTGCTG GCTGCCTTG CTGGCCACG P G A A L A P G G
C P A Q E V 46 CCGGGGGCG GCGCTGGCC CCGGGGGGC TGCCCTGCA CAGGAGGTG A
R G V L T S L P G D S V T L 91 GCGAGAGGT GTGCTGACC AGTCTGCCA
GGAGACAGC GTGACTCTG T C P G G E P E D N A T V H W 136 ACCTGCCCA
GGGGGAGAG CCGGAAGAC AATGCCACT GTTCACTGG V L R K P A E G S H L S R W
A 181 GTTCTCAGG AAGCCAGCT GAAGGCTCC CACCTCAGC AGATGGGCT G V G R R L
L L R S V Q L H D 226 GGCGTGGGA AGGAGGCTG CTGCTGAGG TCGGTGCAG
CTCCATGAC S G N Y S C Y R A G R P A A T 271 TCTGGAAAC TATTCATGC
TACCGGGCC GGCCGCCCG GCTGCAACT V H L L V D V P P E E P Q L S 316
GTGCACTTG CTGGTGGAT GTTCCCCCC GAGGAGCCC CAGCTCTCC C F R K S P L S N
V V C E W G 361 TGCTTCCGG AAGAGCCCA CTCAGCAAC GTTGTTTGT GAGTGGGGT P
R S T P S P T T K A V L L V 406 CCTCGGAGC ACCCCATCT CCGACGACC
AAGGCTGTG CTGTTGGTG R K F Q N S P A E D F Q E P C 451 AGGAAGTTT
CAGAACAGT CCGGCCGAA GACTTCCAG GAGCCGTGC Q Y S Q E S Q K F S C Q L A
V 496 CAGTATTCC CAGGAGTCC CAGAAGTTC TCCTGCCAG TTGGCAGTC P E G D S S
F Y I V S M C V A 541 CCGGAGGGA GACAGCTCT TTCTACATA GTGTCCATG
TGCGTCGCC S S V G S K L S K T Q T F Q G 586 AGTAGTGTC GGGAGCAAG
CTCAGCAAA ACTCAGACC TTTCAGGGT C G I L Q P D P P A N I T V T 631
TGTGGAATC TTGCAGCCT GATCCGCCT GCCAACATC ACAGTCACT A V A R N P R W L
S V T W Q D 676 GCCGTGGCC AGAAACCCC CGCTGGCTC AGTGTCACC TGGCAAGAC P
H S W N S S F Y R L R F E L 721 CCCCACTCC TGGAACTCA TCTTTCTAC
AGACTACGG TTTGAGCTC R Y R A E R S K T F T T W M V 766 AGATATCGA
GCTGAACGG TCAAAGACA TTCACAACA TGGATGGTC K D L Q H H C V I H D A W S
G 811 AAGGACCTC CAGCATCAC TGTGTCATC CACGACGCC TGGAGCGGC L R H V V Q
L R A Q E E F G Q 856 CTGAGGCAC GTGGTGCAG CTTCGTGCC CAGGAGGAG
TTCGGGCAA G E W S E W S P E A M G T P W 901 GGCGAGTGG AGCGAGTGG
AGCCCGGAG GCCATGGGC ACGCCTTGG T E S R S P P A E N E V S T P 946
ACAGAATCC AGGAGTCCT CCAGCTGAG AACGAGGTG TCCACCCCC T Q A P T T N K D
D D N I L S 991 ACGCAGGCA CCTACTACT AATAAAGAT GATGATAAT ATTCTCTCC G
D S A A A T S L P V Q D S S 1036 GGAGATTCT GCAAATGCG ACAAGCCTC
CCAGTGCAA GATTCTTCT S V P L P T F L V A G G S L A 1081 TCGGTACCA
CTGCCCACA TTCCTGGTT GCTGGAGGG AGCCTGGCG F G T L L C I A I V L R F K
K 1126 TTCGGAACG CTCCTGTGC ATTGCCATT GTTCTGAGG TTCAAGAAG T W K L R
A L K E G K T S M H 1171 ACGTGGAAG CTGCGGGCT CTGAAGGAA GGCAAGACA
AGCATGCAC P P Y S L G Q L V P E R P R P 1216 CCGCCGTAT TCTTTGGGG
CAGCTGGTC CCAGAGAGG CCGCGACCC T P V L V P L I S P P V S P S 1261
ACCCCAGTG CTTGTTCCT CTCATCTCC CCACCAGTG TCCCCCAGT S L G S D N T S S
H N R P D A 1306 AGCCTGGGG TCTGACAAC ACCTCGAGC CACAACCGA CCAGATGCC
R D P R S P Y D I S N T D Y F 1351 AGGGACCCA CGGAGCCCT TACGACATC
AGCAATACA GACTACTTC F P R * 1396 TTCCCCAGA TAG
TABLE-US-00003 TABLE 3 Alignment of human vs. cyno gp80. The
nucleotide sequences for human (Genbank Accession BC132684) (SEQ ID
NO: 12) and cyno gp80 (SEQ ID NO: 1) are aligned and are 97%
identical (divergent residues shown in bold). The consensus
sequence (SEQ ID NO: 13) is shown under the aligned human and cyno
gp80 sequences. cyno gp80 (1)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCTTGCTGGCCACGCCGGG human gp80 (1)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCCTGCTGGCCGCGCCGGG cyno gp80 (51)
GGCGGCGCTGGCCCCGGGGGGCTGCCCTGCACAGGAGGTGGCGAGAGGTG human gp80 (51)
AGCGGCGCTGGCCCCAAGGCGCTGCCCTGCGCAGGAGGTGGCGAGAGGCG cyno gp80 (101)
TGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCAGGGGGA human gp80 (101)
TGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCGGGGGTA cyno gp80 (151)
GAGCCGGAAGACAATGCCACTGTTCACTGGGTTCTCAGGAAGCCAGCTGA human gp80 (151)
GAGCCGGAAGACAATGCCACTGTTCACTGGGTGCTCAGGAAGCCGGCTGC cyno gp80 (201)
AGGCTCCCACCTCAGCAGATGGGCTGGCGTGGGAAGGAGGCTGCTGCTGA human gp80 (201)
AGGCTCCCACCCCAGCAGATGGGCTGGCATGGGAAGGAGGCTGCTGCTGA cyno gp80 (251)
GGTCGGTGCAGCTCCATGACTCTGGAAACTATTCATGCTACCGGGCCGGC human gp80 (251)
GGTCGGTGCAGCTCCACGACTCTGGAAACTATTCATGCTACCGGGCCGGC cyno gp80 (301)
CGCCCGGCTGCAACTGTGCACTTGCTGGTGGATGTTCCCCCCGAGGAGCC human gp80 (301)
CGCCCAGCTGGGACTGTGCACTTGCTGGTGGATGTTCCCCCCGAGGAGCC cyno gp80 (351)
CCAGCTCTCCTGCTTCCGGAAGAGCCCACTCAGCAACGTTGTTTGTGAGT human gp80 (351)
CCAGCTCTCCTGCTTCCGGAAGAGCCCCCTCAGCAATGTTGTTTGTGAGT cyno gp80 (401)
GGGGTCCTCGGAGCACCCCATCTCCGACGACCAAGGCTGTGCTGTTGGTG human gp80 (401)
GGGGTCCTCGGAGCACCCCATCCCTGACGACAAAGGCTGTGCTCTTGGTG cyno gp80 (451)
AGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCCAGGAGCCGTGCCAGTA human gp80 (451)
AGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCCAGGAGCCGTGCCAGTA cyno gp80 (501)
TTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTGGCAGTCCCGGAGGGAG human gp80 (501)
TTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTAGCAGTCCCGGAGGGAG cyno gp80 (551)
ACAGCTCTTTCTACATAGTGTCCATGTGCGTCGCCAGTAGTGTCGGGAGC human gp80 (551)
ACAGCTCTTTCTACATAGTGTCCATGTGCGTCGCCAGTAGTGTCGGGAGC cyno gp80 (601)
AAGCTCAGCAAAACTCAGACCTTTCAGGGTTGTGGAATCTTGCAGCCTGA human gp80 (601)
AAGTTCAGCAAAACTCAAACCTTTCAGGGTTGTGGAATCTTGCAGCCTGA cyno gp80 (651)
TCCGCCTGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGC human gp80 (651)
TCCGCCTGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGC cyno gp80 (701)
TCAGTGTCACCTGGCAAGACCCCCACTCCTGGAACTCATCTTTCTACAGA human gp80 (701)
TCAGTGTCACCTGGCAAGACCCCCACTCCTGGAACTCATCTTTCTACAGA cyno gp80 (751)
CTACGGTTTGAGCTCAGATATCGAGCTGAACGGTCAAAGACATTCACAAC human gp80 (751)
CTACGGTTTGAGCTCAGATATCGGGCTGAACGGTCAAAGACATTCACAAC cyno gp80 (801)
ATGGATGGTCAAGGACCTCCAGCATCACTGTGTCATCCACGACGCCTGGA human gp80 (801)
ATGGATGGTCAAGGACCTCCAGCATCACTGTGTCATCCACGACGCCTGGA cyno gp80 (851)
GCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTCGGGCAA human gp80 (851)
GCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTCGGGCAA cyno gp80 (901)
GGCGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGA human gp80 (901)
GGCGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGA cyno gp80 (951)
ATCCAGGAGTCCTCCAGCTGAGAACGAGGTGTCCACCCCCACGCAGGCAC human gp80 (951)
ATCCAGGAGTCCTCCAGCTGAGAACGAGGTGTCCACCCCCATGCAGGCAC cyno gp80 (1001)
CTACTACTAATAAAGATGATGATAATATTCTCTCCGGAGATTCTGCAAAT human gp80
(1001) TTACTACTAATAAAGACGATGATAATATTCTCTTCAGAGATTCTGCAAAT cyno gp80
(1051) GCGACAAGCCTCCCAGTGCAAGATTCTTCTTCGGTACCACTGCCCACATT human
gp80 (1051) GCGACAAGCCTCCCAGTGCAAGATTCTTCTTCAGTACCACTGCCCACATT cyno
gp80 (1101) CCTGGTTGCTGGAGGGAGCCTGGCGTTCGGAACGCTCCTGTGCATTGCCA
human gp80 (1101)
CCTGGTTGCTGGAGGGAGCCTGGCCTTCGGAACGCTCCTCTGCATTGCCA cyno gp80 (1151)
TTGTTCTGAGGTTCAAGAAGACGTGGAAGCTGCGGGCTCTGAAGGAAGGC human gp80
(1151) TTGTTCTGAGGTTCAAGAAGACGTGGAAGCTGCGGGCTCTGAAGGAAGGC cyno gp80
(1201) AAGACAAGCATGCACCCGCCGTATTCTTTGGGGCAGCTGGTCCCAGAGAG human
gp80 (1201) AAGACAAGCATGCATCCGCCGTACTCTTTGGGGCAGCTGGTCCCGGAGAG cyno
gp80 (1251) GCCGCGACCCACCCCAGTGCTTGTTCCTCTCATCTCCCCACCAGTGTCCC
human gp80 (1251)
GCCTCGACCCACCCCAGTGCTTGTTCCTCTCATCTCCCCACCGGTGTCCC cyno gp80 (1301)
CCAGTAGCCTGGGGTCTGACAACACCTCGAGCCACAACCGACCAGATGCC human gp80
(1301) CCAGCAGCCTGGGGTCTGACAATACCTCGAGCCACAACCGACCAGATGCC cyno gp80
(1351) AGGGACCCACGGAGCCCTTACGACATCAGCAATACAGACTACTTCTTCCC human
gp80 (1351) AGGGACCCACGGAGCCCTTATGACATCAGCAATACAGACTACTTCTTCCC cyno
gp80 (1401) CAGATAG human gp80 (1401) CAGATAG
TABLE-US-00004 TABLE 4 Alignment of human and cyno gp80. The
predicted amino acid sequences for human (SEQ ID NO: 11) and cyno
gp80 (SEQ ID NO: 2) are aligned. The cyno sequence is 97% identical
to the human sequence, respectively (divergent residues shown in
bold). 1 50 cyno gp80 (1)
MLAVGCALLAALLATPGAALAPGGCPAQEVARGVLTSLPGDSVTLTCPGG human gp80 v1
NM_000565 (1) MLAVGCALLAALLAAPGAALAPRRCPAQEVARGVLTSLPGDSVTLTCPGV
Consensus (1) MLAVGCALLAALLA PGAALAP CPAQEVARGVLTSLPGDSVTLTCPG 51
100 cyno gp80 (51)
EPEDNATVHWVLRKPAEGSHLSRWAGVGRRLLLRSVQLHDSGNYSCYRAG human gp80 v1
NM_000565 (51) EPEDNATVHWVLRKPAAGSHPSRWAGMGRRLLLRSVQLHDSGNYSCYRAG
Consensus (51) EPEDNATVHWVLRKPA GSH SRWAGMGRRLLLRSVQLHDSGNYSCYRAG
101 150 cyno gp80 (101)
RPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLV human gp80 v1
NM_000565 (101) RPAGTVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSLTT-
KAVLLV Consensus (101) RPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPS
TTKAVLLV 151 200 cyno gp80 (151)
RKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGS human gp80 v1
NM_000565 (151) RKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYIVSMCV-
ASSVGS Consensus (151)
RKFQNSPAEDFQEPCQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGS 201 250 cyno
gp80 (201) KLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYR human
gp80 v1 NM_000565 (201)
KFSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSW- NSSFYR Consensus
(201) K SKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYR 251 300
cyno gp80 (251) LRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQ
human gp80 v1 NM_000565 (251)
LRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRA- QEEFGQ Consensys
(251) LRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQ 301 350
cyno gp80 (301) GEWSEWSPEAMGTPWTESRSPPAENEVSTPTQAPTTNKDDDNILSGDSAN
human gp80 v1 NM_000565 (301)
GEWSEWSPEAMGTPWTESRSPPAENEVSTPMQALTTNKDDDNIL- FRDSAN Consensus
(301) GEWSEWSPEAMGTPWTESRSPPAENEVSTP QA TTNKDDDNIL DSAN 351 400
cyno gp80 (351) ATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEG
human gp80 v1 NM_000565 (351)
ATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKL- RALKEG Consensus
(351) ATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEG 401 450
cyno gp80 (401) KTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSSHNRPDA
human gp80 v1 NM_000565 (401)
KTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSS- HNRPDA Consensus
(401) KTSMHPPYSLGQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSSHNRPDA 451 469
cyno gp80 (451) RDPRSPYDISNTDYFFPR- human gp80 v1 NM_000565 (451)
RDPRSPYDISNTDYFFPR- Consensus (451) RDPRSPYDISNTDYFFPR SEQ ID NO: 1
(signal sequence single underlined) (transmembrane domain double
underlined)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCTTGCTGGCCACGCCGGGGGCGGCGCTGGCCCCGGGGGGCTG-
CCCTGCAC
AGGAGGTGGCGAGAGGTGTGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCAGGGGGAGAGCCG-
GAAGACAA
TGCCACTGTTCACTGGGTTCTCAGGAAGCCAGCTGAAGGCTCCCACCTCAGCAGATGGGCTGGCGTGGGAAGGA-
GGCTGCTG
CTGAGGTCGGTGCAGCTCCATGACTCTGGAAACTATTCATGCTACCGGGCCGGCCGCCCGGCTGCAACTGTGCA-
CTTGCTGG
TGGATGTTCCCCCCGAGGAGCCCCAGCTCTCCTGCTTCCGGAAGAGCCCACTCAGCAACGTTGTTTGTGAGTGG-
GGTCCTCG
GAGCACCCCATCTCCGACGACCAAGGCTGTGCTGTTGGTGAGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCC-
AGGAGCCG
TGCCAGTATTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTGGCAGTCCCGGAGGGAGACAGCTCTTTCTACAT-
AGTGTCCA
TGTGCGTCGCCAGTAGTGTCGGGAGCAAGCTCAGCAAAACTCAGACCTTTCAGGGTTGTGGAATCTTGCAGCCT-
GATCCGCC
TGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGCTCAGTGTCACCTGGCAAGACCCCCACTCCT-
GGAACTCA
TCTTTCTACAGACTACGGTTTGAGCTCAGATATCGAGCTGAACGGTCAAAGACATTCACAACATGGATGGTCAA-
GGACCTCC
AGCATCACTGTGTCATCCACGACGCCTGGAGCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTC-
GGGCAAGG
CGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGAATCCAGGAGTCCTCCAGCTGAGAACG-
AGGTGTCC
ACCCCCACGCAGGCACCTACTACTAATAAAGATGATGATAATATTCTCTCCGGAGATTCTGCAAATGCGACAAG-
CCTCCCAG
TGCAAGATTCTTCTTCGGTACCACTGCCCACATTCCTGGTTGCTGGAGGGAGCCTGGCGTTCGGAACGCTCCTG-
TGCATTGC
CATTGTTCTGAGGTTCAAGAAGACGTGGAAGCTGCGGGCTCTGAAGGAAGGCAAGACAAGCATGCACCCGCCGT-
ATTCTTTG
GGGCAGCTGGTCCCAGAGAGGCCGCGACCCACCCCAGTGCTTGTTCCTCTCATCTCCCCACCAGTGTCCCCCAG-
TAGCCTGG
GGTCTGACAACACCTCGAGCCACAACCGACCAGATGCCAGGGACCCACGGAGCCCTTACGACATCAGCAATACA-
GACTACTT CTTCCCCAGA SEQ ID NO: 2 (signal sequence single
underlined) (transmembrane domain double underlined)
MLAVGCALLAALLATPGAALAPGGCPAQEVARGVLTSLPGDSVTLTCPGGEPEDNATVHWVLRKPAEGSHLSRW-
AGVGRRLL
LRSVQLHDSGNYSCYRAGRPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLVRKFQNS-
PAEDFQEP
CQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTW-
QDPHSWNS
SFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRS-
PPAENEVS
TPTQAPTTNKDDDNILSGDSANATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEGKT-
SMHPPYSL GQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSSHNRPDARDPRSPYDISNTDYFFPR
SEQ ID NO: 3 (his-tagged cyno gp80) (signal sequence single
underlined) (transmembrane domain double underlined)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCTTGCTGGCCACGCCGGGGGCGGCGCTGGCCCCGGGGGGCTG-
CCCTGCAC
AGGAGGTGGCGAGAGGTGTGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCAGGGGGAGAGCCG-
GAAGACAA
TGCCACTGTTCACTGGGTTCTCAGGAAGCCAGCTGAAGGCTCCCACCTCAGCAGATGGGCTGGCGTGGGAAGGA-
GGCTGCTG
CTGAGGTCGGTGCAGCTCCATGACTCTGGAAACTATTCATGCTACCGGGCCGGCCGCCCGGCTGCAACTGTGCA-
CTTGCTGG
TGGATGTTCCCCCCGAGGAGCCCCAGCTCTCCTGCTTCCGGAAGAGCCCACTCAGCAACGTTGTTTGTGAGTGG-
GGTCCTCG
GAGCACCCCATCTCCGACGACCAAGGCTGTGCTGTTGGTGAGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCC-
AGGAGCCG
TGCCAGTATTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTGGCAGTCCCGGAGGGAGACAGCTCTTTCTACAT-
AGTGTCCA
TGTGCGTCGCCAGTAGTGTCGGGAGCAAGCTCAGCAAAACTCAGACCTTTCAGGGTTGTGGAATCTTGCAGCCT-
GATCCGCC
TGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGCTCAGTGTCACCTGGCAAGACCCCCACTCCT-
GGAACTCA
TCTTTCTACAGACTACGGTTTGAGCTCAGATATCGAGCTGAACGGTCAAAGACATTCACAACATGGATGGTCAA-
GGACCTCC
AGCATCACTGTGTCATCCACGACGCCTGGAGCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTC-
GGGCAAGG
CGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGAATCCAGGAGTCCTCCAGCTGAGAACG-
AGGTGTCC
ACCCCCACGCAGGCACCTACTACTAATAAAGATGATGATAATATTCTCTCCGGAGATTCTGCAAATGCGACAAG-
CCTCCCAG
TGCAAGATTCTTCTTCGGTACCACTGCCCACATTCCTGGTTGCTGGAGGGAGCCTGGCGTTCGGAACGCTCCTG-
TGCATTGC
CATTGTTCTGAGGTTCAAGAAGACGTGGAAGCTGCGGGCTCTGAAGGAAGGCAAGACAAGCATGCACCCGCCGT-
ATTCTTTG
GGGCAGCTGGTCCCAGAGAGGCCGCGACCCACCCCAGTGCTTGTTCCTCTCATCTCCCCACCAGTGTCCCCCAG-
TAGCCTGG
GGTCTGACAACACCTCGAGCCACAACCGACCAGATGCCAGGGACCCACGGAGCCCTTACGACATCAGCAATACA-
GACTACTT CTTCCCCAGAGGATCCCATCACCACCACCATCAC SEQ ID NO: 4
(his-tagged cyno gp80) (signal sequence underlined)
MLAVGCALLAALLATPGAALAPGGCPAQEVARGVLTSLPGDSVTLTCPGGEPEDNATVHWVLRKPAEGSHLSRW-
AGVGRRLL
LRSVQLHDSGNYSCYRAGRPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLVRKFQNS-
PAEDFQEP
CQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTW-
QDPHSWNS
SFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRS-
PPAENEVS TPTQAPTTNKDDDNILGSHHHHHH SEQ ID NO: 5 (extracellular
domain of mature cyno gp80) (signal sequence underlined)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCTTGCTGGCCACGCCGGGGGCGGCGCTGGCCCCGGGGGGCTG-
CCCTGCAC
AGGAGGTGGCGAGAGGTGTGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCAGGGGGAGAGCCG-
GAAGACAA
TGCCACTGTTCACTGGGTTCTCAGGAAGCCAGCTGAAGGCTCCCACCTCAGCAGATGGGCTGGCGTGGGAAGGA-
GGCTGCTG
CTGAGGTCGGTGCAGCTCCATGACTCTGGAAACTATTCATGCTACCGGGCCGGCCGCCCGGCTGCAACTGTGCA-
CTTGCTGG
TGGATGTTCCCCCCGAGGAGCCCCAGCTCTCCTGCTTCCGGAAGAGCCCACTCAGCAACGTTGTTTGTGAGTGG-
GGTCCTCG
GAGCACCCCATCTCCGACGACCAAGGCTGTGCTGTTGGTGAGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCC-
AGGAGCCG
TGCCAGTATTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTGGCAGTCCCGGAGGGAGACAGCTCTTTCTACAT-
AGTGTCCA
TGTGCGTCGCCAGTAGTGTCGGGAGCAAGCTCAGCAAAACTCAGACCTTTCAGGGTTGTGGAATCTTGCAGCCT-
GATCCGCC
TGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGCTCAGTGTCACCTGGCAAGACCCCCACTCCT-
GGAACTCA
TCTTTCTACAGACTACGGTTTGAGCTCAGATATCGAGCTGAACGGTCAAAGACATTCACAACATGGATGGTCAA-
GGACCTCC
AGCATCACTGTGTCATCCACGACGCCTGGAGCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTC-
GGGCAAGG
CGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGAATCCAGGAGTCCTCCAGCTGAGAACG-
AGGTGTCC ACCCCCACGCAGGCACCTACTACTAATAAAGATGATGATAATATTCTC SEQ ID
NO: 6 (extracellular domain of mature cyno gp80) (signal sequence
underlined)
MLAVGCALLAALLATPGAALAPGGCPAQEVARGVLTSLPGDSVTLTCPGGEPEDNATVHWVLRKPAEGSHLSRW-
AGVGRRLL
LRSVQLHDSGNYSCYRAGRPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLVRKFQNS-
PAEDFQEP
CQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTW-
QDPHSWNS
SFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRS-
PPAENEVS TPTQAPTTNKDDDNIL SEQ ID NO: 7 (his-tagged extracellular
domain of mature cyno gp80) (signal sequence underlined)
ATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCTTGCTGGCCACGCCGGGGGCGGCGCTGGCCCCGGGGGGCTG-
CCCTGCAC
AGGAGGTGGCGAGAGGTGTGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCAGGGGGAGAGCCG-
GAAGACAA
TGCCACTGTTCACTGGGTTCTCAGGAAGCCAGCTGAAGGCTCCCACCTCAGCAGATGGGCTGGCGTGGGAAGGA-
GGCTGCTG
CTGAGGTCGGTGCAGCTCCATGACTCTGGAAACTATTCATGCTACCGGGCCGGCCGCCCGGCTGCAACTGTGCA-
CTTGCTGG
TGGATGTTCCCCCCGAGGAGCCCCAGCTCTCCTGCTTCCGGAAGAGCCCACTCAGCAACGTTGTTTGTGAGTGG-
GGTCCTCG
GAGCACCCCATCTCCGACGACCAAGGCTGTGCTGTTGGTGAGGAAGTTTCAGAACAGTCCGGCCGAAGACTTCC-
AGGAGCCG
TGCCAGTATTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTGGCAGTCCCGGAGGGAGACAGCTCTTTCTACAT-
AGTGTCCA
TGTGCGTCGCCAGTAGTGTCGGGAGCAAGCTCAGCAAAACTCAGACCTTTCAGGGTTGTGGAATCTTGCAGCCT-
GATCCGCC
TGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGCTCAGTGTCACCTGGCAAGACCCCCACTCCT-
GGAACTCA
TCTTTCTACAGACTACGGTTTGAGCTCAGATATCGAGCTGAACGGTCAAAGACATTCACAACATGGATGGTCAA-
GGACCTCC
AGCATCACTGTGTCATCCACGACGCCTGGAGCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGAGTTC-
GGGCAAGG
CGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGAATCCAGGAGTCCTCCAGCTGAGAACG-
AGGTGTCC
ACCCCCACGCAGGCACCTACTACTAATAAAGATGATGATAATATTCTCGGATCCCATCACCACCACCATCAC
SEQ ID NO: 8 (his-tagged extracellular domain of mature cyno gp80)
(signal sequence underlined)
MLAVGCALLAALLATPGAALAPGGCPAQEVARGVLTSLPGDSVTLTCPGGEPEDNATVHWVLRKPAEGSHLSRW-
AGVGRRLL
LRSVQLHDSGNYSCYRAGRPAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLVRKFQNS-
PAEDFQEP
CQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTW-
QDPHSWNS
SFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRS-
PPAENEVS TPTQAPTTNKDDDNILGSHHHHHH SEQ ID NO: 10 (mature cyno gp80
without signal sequence)
LAPGGCPAQEVARGVLTSLPGDSVTLTCPGGEPEDNATVHWVLRKPAEGSHLSRWAGVGRRLLLRSVQLHDSGN-
YSCYRAGR
PAATVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSPTTKAVLLVRKFQNSPAEDFQEPCQYSQESQKFS-
CQLAVPEG
DSSFYIVSMCVASSVGSKLSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTWQDPHSWNSSFYRLRFELRY-
RAERSKTF
TTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRSPPAENEVSTPTQAPTTNKD-
DDNILSGD
SANATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEGKTSMHPPYSLGQLVPERPRPT-
PVLVPLIS PPVSPSSLGSDNTSSHNRPDARDPRSPYDISNTDYFFPR SEQ ID NO: 11
(human gp80)
MLAVGCALLAALLAAPGAALAPRRCPAQEVARGVLTSLPGDSVTLTCPGVEPEDNATVHWVLRKPAAGSHPSRW-
AGMGRRLL
LRSVQLHDSGNYSCYRAGRPAGTVHLLVDVPPEEPQLSCFRKSPLSNVVCEWGPRSTPSLTTKAVLLVRKFQNS-
PAEDFQEP
CQYSQESQKFSCQLAVPEGDSSFYIVSMCVASSVGSKFSKTQTFQGCGILQPDPPANITVTAVARNPRWLSVTW-
QDPHSWNS
SFYRLRFELRYRAERSKTFTTWMVKDLQHHCVIHDAWSGLRHVVQLRAQEEFGQGEWSEWSPEAMGTPWTESRS-
PPAENEVS
TPMQALTTNKDDDNILFRDSANATSLPVQDSSSVPLPTFLVAGGSLAFGTLLCIAIVLRFKKTWKLRALKEGKT-
SMHPPYSL GQLVPERPRPTPVLVPLISPPVSPSSLGSDNTSSHNRPDARDPRSPYDISNTDYFFPR
SEQ ID NO: 12 (Homo sapiens)
TGAGTCATGTGCGAGTGGGAAGTCGCACTGACACTGAGCCGGGCCAGAGGGAGAGGAGCCGAGCGCGGCGCGGG-
GCCGAGGG
ACTCGCAGTGTGTGTAGAGAGCCGGGCTCCTGCGGATGGGGGCTGCCCCCGGGGCCTGAGCCCGCCTGCCCGCC-
CACCGCCC
CGCCCCGCCCCTGCCACCCCTGCCGCCCGGTTCCCATTAGCCTGTCCGCCTCTGCGGGACCATGGAGTGGTAGC-
CGAGGAGG
AAGCATGCTGGCCGTCGGCTGCGCGCTGCTGGCTGCCCTGCTGGCCGCGCCGGGAGCGGCGCTGGCCCCAAGGC-
GCTGCCCT
GCGCAGGAGGTGGCGAGAGGCGTGCTGACCAGTCTGCCAGGAGACAGCGTGACTCTGACCTGCCCGGGGGTAGA-
GCCGGAAG
ACAATGCCACTGTTCACTGGGTGCTCAGGAAGCCGGCTGCAGGCTCCCACCCCAGCAGATGGGCTGGCATGGGA-
AGGAGGCT
GCTGCTGAGGTCGGTGCAGCTCCACGACTCTGGAAACTATTCATGCTACCGGGCCGGCCGCCCAGCTGGGACTG-
TGCACTTG
CTGGTGGATGTTCCCCCCGAGGAGCCCCAGCTCTCCTGCTTCCGGAAGAGCCCCCTCAGCAATGTTGTTTGTGA-
GTGGGGTC
CTCGGAGCACCCCATCCCTGACGACAAAGGCTGTGCTCTTGGTGAGGAAGTTTCAGAACAGTCCGGCCGAAGAC-
TTCCAGGA
GCCGTGCCAGTATTCCCAGGAGTCCCAGAAGTTCTCCTGCCAGTTAGCAGTCCCGGAGGGAGACAGCTCTTTCT-
ACATAGTG
TCCATGTGCGTCGCCAGTAGTGTCGGGAGCAAGTTCAGCAAAACTCAAACCTTTCAGGGTTGTGGAATCTTGCA-
GCCTGATC
CGCCTGCCAACATCACAGTCACTGCCGTGGCCAGAAACCCCCGCTGGCTCAGTGTCACCTGGCAAGACCCCCAC-
TCCTGGAA
CTCATCTTTCTACAGACTACGGTTTGAGCTCAGATATCGGGCTGAACGGTCAAAGACATTCACAACATGGATGG-
TCAAGGAC
CTCCAGCATCACTGTGTCATCCACGACGCCTGGAGCGGCCTGAGGCACGTGGTGCAGCTTCGTGCCCAGGAGGA-
GTTCGGGC
AAGGCGAGTGGAGCGAGTGGAGCCCGGAGGCCATGGGCACGCCTTGGACAGAATCCAGGAGTCCTCCAGCTGAG-
AACGAGGT
GTCCACCCCCATGCAGGCACTTACTACTAATAAAGACGATGATAATATTCTCTTCAGAGATTCTGCAAATGCGA-
CAAGCCTC
CCAGTGCAAGATTCTTCTTCAGTACCACTGCCCACATTCCTGGTTGCTGGAGGGAGCCTGGCCTTCGGAACGCT-
CCTCTGCA
TTGCCATTGTTCTGAGGTTCAAGAAGACGTGGAAGCTGCGGGCTCTGAAGGAAGGCAAGACAAGCATGCATCCG-
CCGTACTC
TTTGGGGCAGCTGGTCCCGGAGAGGCCTCGACCCACCCCAGTGCTTGTTCCTCTCATCTCCCCACCGGTGTCCC-
CCAGCAGC
CTGGGGTCTGACAATACCTCGAGCCACAACCGACCAGATGCCAGGGACCCACGGAGCCCTTATGACATCAGCAA-
TACAGACT
ACTTCTTCCCCAGATAGCTGGCTGGGTGGCACCAGCAGCCTGGACCCTGTGGATGATAAAACACAAACGGGCTC-
AGCA
SEQUENCE LISTINGS
1
1311404DNAMacaca fascicularis 1atgctggccg tcggctgcgc gctgctggct
gccttgctgg ccacgccggg ggcggcgctg 60gccccggggg gctgccctgc acaggaggtg
gcgagaggtg tgctgaccag tctgccagga 120gacagcgtga ctctgacctg
cccaggggga gagccggaag acaatgccac tgttcactgg 180gttctcagga
agccagctga aggctcccac ctcagcagat gggctggcgt gggaaggagg
240ctgctgctga ggtcggtgca gctccatgac tctggaaact attcatgcta
ccgggccggc 300cgcccggctg caactgtgca cttgctggtg gatgttcccc
ccgaggagcc ccagctctcc 360tgcttccgga agagcccact cagcaacgtt
gtttgtgagt ggggtcctcg gagcacccca 420tctccgacga ccaaggctgt
gctgttggtg aggaagtttc agaacagtcc ggccgaagac 480ttccaggagc
cgtgccagta ttcccaggag tcccagaagt tctcctgcca gttggcagtc
540ccggagggag acagctcttt ctacatagtg tccatgtgcg tcgccagtag
tgtcgggagc 600aagctcagca aaactcagac ctttcagggt tgtggaatct
tgcagcctga tccgcctgcc 660aacatcacag tcactgccgt ggccagaaac
ccccgctggc tcagtgtcac ctggcaagac 720ccccactcct ggaactcatc
tttctacaga ctacggtttg agctcagata tcgagctgaa 780cggtcaaaga
cattcacaac atggatggtc aaggacctcc agcatcactg tgtcatccac
840gacgcctgga gcggcctgag gcacgtggtg cagcttcgtg cccaggagga
gttcgggcaa 900ggcgagtgga gcgagtggag cccggaggcc atgggcacgc
cttggacaga atccaggagt 960cctccagctg agaacgaggt gtccaccccc
acgcaggcac ctactactaa taaagatgat 1020gataatattc tctccggaga
ttctgcaaat gcgacaagcc tcccagtgca agattcttct 1080tcggtaccac
tgcccacatt cctggttgct ggagggagcc tggcgttcgg aacgctcctg
1140tgcattgcca ttgttctgag gttcaagaag acgtggaagc tgcgggctct
gaaggaaggc 1200aagacaagca tgcacccgcc gtattctttg gggcagctgg
tcccagagag gccgcgaccc 1260accccagtgc ttgttcctct catctcccca
ccagtgtccc ccagtagcct ggggtctgac 1320aacacctcga gccacaaccg
accagatgcc agggacccac ggagccctta cgacatcagc 1380aatacagact
acttcttccc caga 14042468PRTMacaca fascicularis 2Met Leu Ala Val Gly
Cys Ala Leu Leu Ala Ala Leu Leu Ala Thr Pro 1 5 10 15Gly Ala Ala
Leu Ala Pro Gly Gly Cys Pro Ala Gln Glu Val Ala Arg 20 25 30Gly Val
Leu Thr Ser Leu Pro Gly Asp Ser Val Thr Leu Thr Cys Pro 35 40 45Gly
Gly Glu Pro Glu Asp Asn Ala Thr Val His Trp Val Leu Arg Lys 50 55
60Pro Ala Glu Gly Ser His Leu Ser Arg Trp Ala Gly Val Gly Arg Arg65
70 75 80Leu Leu Leu Arg Ser Val Gln Leu His Asp Ser Gly Asn Tyr Ser
Cys 85 90 95Tyr Arg Ala Gly Arg Pro Ala Ala Thr Val His Leu Leu Val
Asp Val 100 105 110Pro Pro Glu Glu Pro Gln Leu Ser Cys Phe Arg Lys
Ser Pro Leu Ser 115 120 125Asn Val Val Cys Glu Trp Gly Pro Arg Ser
Thr Pro Ser Pro Thr Thr 130 135 140Lys Ala Val Leu Leu Val Arg Lys
Phe Gln Asn Ser Pro Ala Glu Asp145 150 155 160Phe Gln Glu Pro Cys
Gln Tyr Ser Gln Glu Ser Gln Lys Phe Ser Cys 165 170 175Gln Leu Ala
Val Pro Glu Gly Asp Ser Ser Phe Tyr Ile Val Ser Met 180 185 190Cys
Val Ala Ser Ser Val Gly Ser Lys Leu Ser Lys Thr Gln Thr Phe 195 200
205Gln Gly Cys Gly Ile Leu Gln Pro Asp Pro Pro Ala Asn Ile Thr Val
210 215 220Thr Ala Val Ala Arg Asn Pro Arg Trp Leu Ser Val Thr Trp
Gln Asp225 230 235 240Pro His Ser Trp Asn Ser Ser Phe Tyr Arg Leu
Arg Phe Glu Leu Arg 245 250 255Tyr Arg Ala Glu Arg Ser Lys Thr Phe
Thr Thr Trp Met Val Lys Asp 260 265 270Leu Gln His His Cys Val Ile
His Asp Ala Trp Ser Gly Leu Arg His 275 280 285Val Val Gln Leu Arg
Ala Gln Glu Glu Phe Gly Gln Gly Glu Trp Ser 290 295 300Glu Trp Ser
Pro Glu Ala Met Gly Thr Pro Trp Thr Glu Ser Arg Ser305 310 315
320Pro Pro Ala Glu Asn Glu Val Ser Thr Pro Thr Gln Ala Pro Thr Thr
325 330 335Asn Lys Asp Asp Asp Asn Ile Leu Ser Gly Asp Ser Ala Asn
Ala Thr 340 345 350Ser Leu Pro Val Gln Asp Ser Ser Ser Val Pro Leu
Pro Thr Phe Leu 355 360 365Val Ala Gly Gly Ser Leu Ala Phe Gly Thr
Leu Leu Cys Ile Ala Ile 370 375 380Val Leu Arg Phe Lys Lys Thr Trp
Lys Leu Arg Ala Leu Lys Glu Gly385 390 395 400Lys Thr Ser Met His
Pro Pro Tyr Ser Leu Gly Gln Leu Val Pro Glu 405 410 415Arg Pro Arg
Pro Thr Pro Val Leu Val Pro Leu Ile Ser Pro Pro Val 420 425 430Ser
Pro Ser Ser Leu Gly Ser Asp Asn Thr Ser Ser His Asn Arg Pro 435 440
445Asp Ala Arg Asp Pro Arg Ser Pro Tyr Asp Ile Ser Asn Thr Asp Tyr
450 455 460Phe Phe Pro Arg46531428DNAMacaca fascicularis
3atgctggccg tcggctgcgc gctgctggct gccttgctgg ccacgccggg ggcggcgctg
60gccccggggg gctgccctgc acaggaggtg gcgagaggtg tgctgaccag tctgccagga
120gacagcgtga ctctgacctg cccaggggga gagccggaag acaatgccac
tgttcactgg 180gttctcagga agccagctga aggctcccac ctcagcagat
gggctggcgt gggaaggagg 240ctgctgctga ggtcggtgca gctccatgac
tctggaaact attcatgcta ccgggccggc 300cgcccggctg caactgtgca
cttgctggtg gatgttcccc ccgaggagcc ccagctctcc 360tgcttccgga
agagcccact cagcaacgtt gtttgtgagt ggggtcctcg gagcacccca
420tctccgacga ccaaggctgt gctgttggtg aggaagtttc agaacagtcc
ggccgaagac 480ttccaggagc cgtgccagta ttcccaggag tcccagaagt
tctcctgcca gttggcagtc 540ccggagggag acagctcttt ctacatagtg
tccatgtgcg tcgccagtag tgtcgggagc 600aagctcagca aaactcagac
ctttcagggt tgtggaatct tgcagcctga tccgcctgcc 660aacatcacag
tcactgccgt ggccagaaac ccccgctggc tcagtgtcac ctggcaagac
720ccccactcct ggaactcatc tttctacaga ctacggtttg agctcagata
tcgagctgaa 780cggtcaaaga cattcacaac atggatggtc aaggacctcc
agcatcactg tgtcatccac 840gacgcctgga gcggcctgag gcacgtggtg
cagcttcgtg cccaggagga gttcgggcaa 900ggcgagtgga gcgagtggag
cccggaggcc atgggcacgc cttggacaga atccaggagt 960cctccagctg
agaacgaggt gtccaccccc acgcaggcac ctactactaa taaagatgat
1020gataatattc tctccggaga ttctgcaaat gcgacaagcc tcccagtgca
agattcttct 1080tcggtaccac tgcccacatt cctggttgct ggagggagcc
tggcgttcgg aacgctcctg 1140tgcattgcca ttgttctgag gttcaagaag
acgtggaagc tgcgggctct gaaggaaggc 1200aagacaagca tgcacccgcc
gtattctttg gggcagctgg tcccagagag gccgcgaccc 1260accccagtgc
ttgttcctct catctcccca ccagtgtccc ccagtagcct ggggtctgac
1320aacacctcga gccacaaccg accagatgcc agggacccac ggagccctta
cgacatcagc 1380aatacagact acttcttccc cagaggatcc catcaccacc accatcac
14284352PRTMacaca fascicularis 4Met Leu Ala Val Gly Cys Ala Leu Leu
Ala Ala Leu Leu Ala Thr Pro 1 5 10 15Gly Ala Ala Leu Ala Pro Gly
Gly Cys Pro Ala Gln Glu Val Ala Arg 20 25 30Gly Val Leu Thr Ser Leu
Pro Gly Asp Ser Val Thr Leu Thr Cys Pro 35 40 45Gly Gly Glu Pro Glu
Asp Asn Ala Thr Val His Trp Val Leu Arg Lys 50 55 60Pro Ala Glu Gly
Ser His Leu Ser Arg Trp Ala Gly Val Gly Arg Arg65 70 75 80Leu Leu
Leu Arg Ser Val Gln Leu His Asp Ser Gly Asn Tyr Ser Cys 85 90 95Tyr
Arg Ala Gly Arg Pro Ala Ala Thr Val His Leu Leu Val Asp Val 100 105
110Pro Pro Glu Glu Pro Gln Leu Ser Cys Phe Arg Lys Ser Pro Leu Ser
115 120 125Asn Val Val Cys Glu Trp Gly Pro Arg Ser Thr Pro Ser Pro
Thr Thr 130 135 140Lys Ala Val Leu Leu Val Arg Lys Phe Gln Asn Ser
Pro Ala Glu Asp145 150 155 160Phe Gln Glu Pro Cys Gln Tyr Ser Gln
Glu Ser Gln Lys Phe Ser Cys 165 170 175Gln Leu Ala Val Pro Glu Gly
Asp Ser Ser Phe Tyr Ile Val Ser Met 180 185 190Cys Val Ala Ser Ser
Val Gly Ser Lys Leu Ser Lys Thr Gln Thr Phe 195 200 205Gln Gly Cys
Gly Ile Leu Gln Pro Asp Pro Pro Ala Asn Ile Thr Val 210 215 220Thr
Ala Val Ala Arg Asn Pro Arg Trp Leu Ser Val Thr Trp Gln Asp225 230
235 240Pro His Ser Trp Asn Ser Ser Phe Tyr Arg Leu Arg Phe Glu Leu
Arg 245 250 255Tyr Arg Ala Glu Arg Ser Lys Thr Phe Thr Thr Trp Met
Val Lys Asp 260 265 270Leu Gln His His Cys Val Ile His Asp Ala Trp
Ser Gly Leu Arg His 275 280 285Val Val Gln Leu Arg Ala Gln Glu Glu
Phe Gly Gln Gly Glu Trp Ser 290 295 300Glu Trp Ser Pro Glu Ala Met
Gly Thr Pro Trp Thr Glu Ser Arg Ser305 310 315 320Pro Pro Ala Glu
Asn Glu Val Ser Thr Pro Thr Gln Ala Pro Thr Thr 325 330 335Asn Lys
Asp Asp Asp Asn Ile Leu Gly Ser His His His His His His 340 345
35051032DNAMacaca fascicularis 5atgctggccg tcggctgcgc gctgctggct
gccttgctgg ccacgccggg ggcggcgctg 60gccccggggg gctgccctgc acaggaggtg
gcgagaggtg tgctgaccag tctgccagga 120gacagcgtga ctctgacctg
cccaggggga gagccggaag acaatgccac tgttcactgg 180gttctcagga
agccagctga aggctcccac ctcagcagat gggctggcgt gggaaggagg
240ctgctgctga ggtcggtgca gctccatgac tctggaaact attcatgcta
ccgggccggc 300cgcccggctg caactgtgca cttgctggtg gatgttcccc
ccgaggagcc ccagctctcc 360tgcttccgga agagcccact cagcaacgtt
gtttgtgagt ggggtcctcg gagcacccca 420tctccgacga ccaaggctgt
gctgttggtg aggaagtttc agaacagtcc ggccgaagac 480ttccaggagc
cgtgccagta ttcccaggag tcccagaagt tctcctgcca gttggcagtc
540ccggagggag acagctcttt ctacatagtg tccatgtgcg tcgccagtag
tgtcgggagc 600aagctcagca aaactcagac ctttcagggt tgtggaatct
tgcagcctga tccgcctgcc 660aacatcacag tcactgccgt ggccagaaac
ccccgctggc tcagtgtcac ctggcaagac 720ccccactcct ggaactcatc
tttctacaga ctacggtttg agctcagata tcgagctgaa 780cggtcaaaga
cattcacaac atggatggtc aaggacctcc agcatcactg tgtcatccac
840gacgcctgga gcggcctgag gcacgtggtg cagcttcgtg cccaggagga
gttcgggcaa 900ggcgagtgga gcgagtggag cccggaggcc atgggcacgc
cttggacaga atccaggagt 960cctccagctg agaacgaggt gtccaccccc
acgcaggcac ctactactaa taaagatgat 1020gataatattc tc
10326344PRTMacaca fascicularis 6Met Leu Ala Val Gly Cys Ala Leu Leu
Ala Ala Leu Leu Ala Thr Pro 1 5 10 15Gly Ala Ala Leu Ala Pro Gly
Gly Cys Pro Ala Gln Glu Val Ala Arg 20 25 30Gly Val Leu Thr Ser Leu
Pro Gly Asp Ser Val Thr Leu Thr Cys Pro 35 40 45Gly Gly Glu Pro Glu
Asp Asn Ala Thr Val His Trp Val Leu Arg Lys 50 55 60Pro Ala Glu Gly
Ser His Leu Ser Arg Trp Ala Gly Val Gly Arg Arg65 70 75 80Leu Leu
Leu Arg Ser Val Gln Leu His Asp Ser Gly Asn Tyr Ser Cys 85 90 95Tyr
Arg Ala Gly Arg Pro Ala Ala Thr Val His Leu Leu Val Asp Val 100 105
110Pro Pro Glu Glu Pro Gln Leu Ser Cys Phe Arg Lys Ser Pro Leu Ser
115 120 125Asn Val Val Cys Glu Trp Gly Pro Arg Ser Thr Pro Ser Pro
Thr Thr 130 135 140Lys Ala Val Leu Leu Val Arg Lys Phe Gln Asn Ser
Pro Ala Glu Asp145 150 155 160Phe Gln Glu Pro Cys Gln Tyr Ser Gln
Glu Ser Gln Lys Phe Ser Cys 165 170 175Gln Leu Ala Val Pro Glu Gly
Asp Ser Ser Phe Tyr Ile Val Ser Met 180 185 190Cys Val Ala Ser Ser
Val Gly Ser Lys Leu Ser Lys Thr Gln Thr Phe 195 200 205Gln Gly Cys
Gly Ile Leu Gln Pro Asp Pro Pro Ala Asn Ile Thr Val 210 215 220Thr
Ala Val Ala Arg Asn Pro Arg Trp Leu Ser Val Thr Trp Gln Asp225 230
235 240Pro His Ser Trp Asn Ser Ser Phe Tyr Arg Leu Arg Phe Glu Leu
Arg 245 250 255Tyr Arg Ala Glu Arg Ser Lys Thr Phe Thr Thr Trp Met
Val Lys Asp 260 265 270Leu Gln His His Cys Val Ile His Asp Ala Trp
Ser Gly Leu Arg His 275 280 285Val Val Gln Leu Arg Ala Gln Glu Glu
Phe Gly Gln Gly Glu Trp Ser 290 295 300Glu Trp Ser Pro Glu Ala Met
Gly Thr Pro Trp Thr Glu Ser Arg Ser305 310 315 320Pro Pro Ala Glu
Asn Glu Val Ser Thr Pro Thr Gln Ala Pro Thr Thr 325 330 335Asn Lys
Asp Asp Asp Asn Ile Leu 34071056DNAMacaca fascicularis 7atgctggccg
tcggctgcgc gctgctggct gccttgctgg ccacgccggg ggcggcgctg 60gccccggggg
gctgccctgc acaggaggtg gcgagaggtg tgctgaccag tctgccagga
120gacagcgtga ctctgacctg cccaggggga gagccggaag acaatgccac
tgttcactgg 180gttctcagga agccagctga aggctcccac ctcagcagat
gggctggcgt gggaaggagg 240ctgctgctga ggtcggtgca gctccatgac
tctggaaact attcatgcta ccgggccggc 300cgcccggctg caactgtgca
cttgctggtg gatgttcccc ccgaggagcc ccagctctcc 360tgcttccgga
agagcccact cagcaacgtt gtttgtgagt ggggtcctcg gagcacccca
420tctccgacga ccaaggctgt gctgttggtg aggaagtttc agaacagtcc
ggccgaagac 480ttccaggagc cgtgccagta ttcccaggag tcccagaagt
tctcctgcca gttggcagtc 540ccggagggag acagctcttt ctacatagtg
tccatgtgcg tcgccagtag tgtcgggagc 600aagctcagca aaactcagac
ctttcagggt tgtggaatct tgcagcctga tccgcctgcc 660aacatcacag
tcactgccgt ggccagaaac ccccgctggc tcagtgtcac ctggcaagac
720ccccactcct ggaactcatc tttctacaga ctacggtttg agctcagata
tcgagctgaa 780cggtcaaaga cattcacaac atggatggtc aaggacctcc
agcatcactg tgtcatccac 840gacgcctgga gcggcctgag gcacgtggtg
cagcttcgtg cccaggagga gttcgggcaa 900ggcgagtgga gcgagtggag
cccggaggcc atgggcacgc cttggacaga atccaggagt 960cctccagctg
agaacgaggt gtccaccccc acgcaggcac ctactactaa taaagatgat
1020gataatattc tcggatccca tcaccaccac catcac 10568352PRTMacaca
fascicularis 8Met Leu Ala Val Gly Cys Ala Leu Leu Ala Ala Leu Leu
Ala Thr Pro 1 5 10 15Gly Ala Ala Leu Ala Pro Gly Gly Cys Pro Ala
Gln Glu Val Ala Arg 20 25 30Gly Val Leu Thr Ser Leu Pro Gly Asp Ser
Val Thr Leu Thr Cys Pro 35 40 45Gly Gly Glu Pro Glu Asp Asn Ala Thr
Val His Trp Val Leu Arg Lys 50 55 60Pro Ala Glu Gly Ser His Leu Ser
Arg Trp Ala Gly Val Gly Arg Arg65 70 75 80Leu Leu Leu Arg Ser Val
Gln Leu His Asp Ser Gly Asn Tyr Ser Cys 85 90 95Tyr Arg Ala Gly Arg
Pro Ala Ala Thr Val His Leu Leu Val Asp Val 100 105 110Pro Pro Glu
Glu Pro Gln Leu Ser Cys Phe Arg Lys Ser Pro Leu Ser 115 120 125Asn
Val Val Cys Glu Trp Gly Pro Arg Ser Thr Pro Ser Pro Thr Thr 130 135
140Lys Ala Val Leu Leu Val Arg Lys Phe Gln Asn Ser Pro Ala Glu
Asp145 150 155 160Phe Gln Glu Pro Cys Gln Tyr Ser Gln Glu Ser Gln
Lys Phe Ser Cys 165 170 175Gln Leu Ala Val Pro Glu Gly Asp Ser Ser
Phe Tyr Ile Val Ser Met 180 185 190Cys Val Ala Ser Ser Val Gly Ser
Lys Leu Ser Lys Thr Gln Thr Phe 195 200 205Gln Gly Cys Gly Ile Leu
Gln Pro Asp Pro Pro Ala Asn Ile Thr Val 210 215 220Thr Ala Val Ala
Arg Asn Pro Arg Trp Leu Ser Val Thr Trp Gln Asp225 230 235 240Pro
His Ser Trp Asn Ser Ser Phe Tyr Arg Leu Arg Phe Glu Leu Arg 245 250
255Tyr Arg Ala Glu Arg Ser Lys Thr Phe Thr Thr Trp Met Val Lys Asp
260 265 270Leu Gln His His Cys Val Ile His Asp Ala Trp Ser Gly Leu
Arg His 275 280 285Val Val Gln Leu Arg Ala Gln Glu Glu Phe Gly Gln
Gly Glu Trp Ser 290 295 300Glu Trp Ser Pro Glu Ala Met Gly Thr Pro
Trp Thr Glu Ser Arg Ser305 310 315 320Pro Pro Ala Glu Asn Glu Val
Ser Thr Pro Thr Gln Ala Pro Thr Thr 325 330 335Asn Lys Asp Asp Asp
Asn Ile Leu Gly Ser His His His His His His 340 345
35095920DNAMacaca mulatta 9ggcggtcccc tgttctcccc gctcaggtgc
ggcgctgtgg caggaagcca ccccctcggt 60cggccggtgc gcggggctgt tgcgccatcc
gctccagctt tcgtaaccgc accctgggac 120ggcccagaga cgctccagcg
cgagttcctc aaatgttttc ctgtgttgcc aggaccgtcc 180gccgctctga
gtcatgtgcg agtgggaagt ctcactgaca ctgagaccgg ccagtgggag
240cggagccgag cgcggcgcgg ggctgaggga ctcgcagtgt atatagagcg
ccgggctcct 300gcgatggggg ctgcccccgg agactgagcc agcctgcccg
cccaccgccc cgcccctgcc 360gcccggttcc caccagcctg tccgcctctg
cgggaccatg gagcggtagc cgaggaggaa 420gcatgctggc cgtcggctgc
gcgctgctgg ctgccctgct ggccgcgccg ggggcggcgc 480tggccccggg
gggctgccct gcgcaggagg tggcgagagg tgtgctgacc agtctgccag
540gagacagcgt gactctgacc tgcccagggg gagagccgga
agacaatgcc actgttcact 600gggttctcag gaagccagct gtaggctccc
acctcagcag atgggctggc gtgggaagga 660ggctgctgct gaggtcggtg
cagctccatg actctggaaa ctattcatgc taccgggccg 720gccgcccggc
tggaactgtg cacttgctgg tggatgttcc ccccgaggag ccccagctct
780cctgcttccg gaagagcccc ctcagcaacg ttgtttgtga gtggggtcct
cggagcaccc 840catctccgac gaccaaggct gtgctgttgg tgaggaagtt
tcagaacagt ccggccgaag 900acttccagga gccgtgccag tattcccagg
agtcccagaa gttctcctgc cagttggcag 960tcccggaggg agacagctct
ttctacatag tgtccatgtg cgtcgccagt agtgtcggga 1020gcaagctcag
caaaactcag acctttcagg gttgtggaat cttgcagcct gatccgcctg
1080ccaacatcac agtcactgcc gtggccagaa acccccgctg gctcagtgtc
acctggcaag 1140acccccactc ctggaactca tctttctaca gactacggtt
tgagctcaga tatcgagctg 1200aacggtcaaa gacattcaca acatggatgg
tcaaggacct ccagcatcac tgtgtcatcc 1260acgacgcctg gagcggcctg
aggcacgtgg tgcagcttcg tgcccaggag gagttcgggc 1320aaggcgagtg
gagcgagtgg agcccggagg ccatgggcac gccttggaca gaatccagga
1380gtcctccagc tgagaacgag gtgtccaccc ccacgcaggc acctactact
aataaagatg 1440atgataatat tctctccaga gattctgcaa atgcgacaag
cctcccagtg caagattctt 1500cttcggtacc actgcccaca ttcctggttg
ctggagggag cctggcgttc ggaacgctcc 1560tgtgcattgc cattgttctg
aggttcaaga agacgtggaa gctgcgggct ctgaaggaag 1620gcaagacaag
catgcacccg ccgtattctt tggggcagct ggtcccagag aggccgcgac
1680ccaccccagt gcttgttcct ctcatctccc caccagtgtc ccccagcagc
ctggggtctg 1740acaacacctc gagccacaac cgaccagatg ccagggaccc
acggagccct tacgacatca 1800gcaatacaga ctacttcttc cccagatagt
tggccagggg gcactagcag gctggaccct 1860gtggatgaca gagcacaaac
gggctcagca aaggatgctt cttactgcca tgccagctta 1920tctcagggga
gtggggcctt tggcttcacg gaagagcctt gcggaaggtt cgacaccagg
1980ggaaaatcag cctgctccag ctgttcagct ggttgagatt tcaaacctcc
ctttccaaat 2040gtctggctta aaggggttag agtgaacttg ggccactgtg
aagagaagca tgtcaagact 2100ctttggacat tcaacacgga cactcaaaag
ctgggcagct tggtgggggc ctcagtgtgg 2160agaagcggct ggcagcccac
cccccaacac ctctgcacaa gctgcgccct caggcagggg 2220gggcggattt
ccagccaaag cctccttcag ccgccacgct cctggcccac tgcatcattt
2280catcttccag ctcaaactcc taaaacccaa gtgcctttgc aaattctgtt
tttctgagcc 2340tggggacggc ttttacttaa accgccaagg ctggggaaag
aagctctctc ctccctttct 2400tccccacagt tgaaaaacag ctgagggtgg
gtgggtgaat aatacagtat ctcagggcct 2460ggtcgttttc aacggaatta
taattagttc ctcattagca ttttgctaaa tgtgaatgat 2520aatcctaggc
atttgctgaa tacagaggca actgcattgg ctttgggttg caggacctca
2580ggtgagaagc agaggaagga gaggagaggg gcacagggtc tccaccatcc
cctgtagagt 2640gggagctgcg cgggggatca cagcctctga aaaccaatgt
tctctgttct ccacctccca 2700caaaggagag ctggcagcag ggagggcttc
tgccagtgct gagatcaaaa ctgttttact 2760gcagctttgt ttgttgtcag
ccgaacctgg gtaactaggg aagataatat taaggaagac 2820aatgtgaaaa
gaaaaatgag cccggcaaga atgcatttta acttggtttt taaaaaactg
2880ctgactgttt tctcttgaga gggtggaata tccaatatgc gctgtgtcag
catagaagta 2940acttacttag gtgtggggga agcaccataa ctttgtttag
cccaaaacta agtcaagtga 3000aaaaggagga agagaaataa tatttttcct
gccaggcatg gtggttcacg cctgtaatcc 3060cagcactctg ggaggtcgag
gcgggacgat cacttgagtc caggagtttg agaccagcct 3120gggcaatgtg
gtaaaacctc atctcgacaa aaagcataaa aattagccag gtatggtaga
3180gtgcacctga agtcccagtt agttgggagg ctgaggtggg aggatctctt
gagcctggga 3240ggtcaaggct gcagtgagcc gagattgcac cactgcactc
cagcctgggt gactgagcaa 3300gtgagaccct gtctcaaaaa agaaaaggaa
aaagaaaaga aaaaatattt tccctgttag 3360agaagagatt gtggtttcat
tgtgtatttt gtttttgtct taaaaagtgg aaaaatagcc 3420tgcctcttct
ctactctagg gaaaaaccag tgtgtgacta ctcccccagg cggttatgga
3480gagggcgtcc ggtccctgtc ccagtgctga gaagggagcc tcccacgact
acccggcagg 3540gtcctagaaa ttccccaccc tgaaagccct gagccttctg
ctatcaaagg ggcaggtaaa 3600aatcccattt aaaaaaaatc ccttacctcg
gtgccttcct ctttttattt agctccttga 3660gttgattcag ctctgcaaga
attgaagcag aactaaatgt ctaattgtaa caccgtgatt 3720aaccacttca
gctgactttt ctgcccgagc tttgaaaatt cagtggtgtt agtggttacc
3780cagttagctc tcaagttatc agggtactcc acagcgggga tataccagac
cacaaaacct 3840ttctaatact ctaccctctt agaaaaacag ccaccatcac
cagacaggtg caaaaggagg 3900aaagtgacca tgttttgttt accgttttcc
aggtttaagc tgttactgtc ttcagcaagc 3960cgtgcttttc attgctgggt
ttgtctgtag attttagacc ctattgctgc ttgaggcacc 4020tcatcttaag
ttggcaaaaa ggcaggacgg ctgggtgtgg tggctcacgc ctgtaatcct
4080agcactttgg gaggccgagg tgggaggatt gcttgagctc aggaatttga
gaccaacctg 4140ggtaacatag tgagatacca tctctattat aaacaataac
atttaaggaa aaaaaaaggc 4200aggcaggtgg ttatggtggt tccctcccat
cctgctgcat aaagtttctg agacttgaga 4260acagcaaaaa tgctgttaaa
gggaaatatt aagaatgaga atctgcatga agggtgatta 4320tgtgcccaca
gttaattctt tataccgttt tacccacatg tggtattacc gaagccgggc
4380agaaccatgc tagcggaaga tatgaaattc agatagctca ttattgccaa
gagctaggca 4440gctttgatct ccaaattgtt attgctttca tttttattgt
aatggaactg cttttttttt 4500tttttttttt ttttttgctt ttttttcttt
gttttgtttt tgtagtgaag agggtttttt 4560tccctttatt tttcataagc
tactgtaaat gaagaaaaag tgtcttctct gggctgtagg 4620cctggctcag
tgtacacagg tatacatcct aagctctctc tgttctctaa tttgtggtga
4680ctgaatatgt gtcgcaatcc acggggcatt tctacctgta tttctgcagc
acccccactg 4740ccttgagtcc ccagcagtgc tgttatttgc ctaatacctg
tagccatctg ccacacagcc 4800agacatgaaa cgctgggaca gagaccattt
agattaaata caacagctta tcttgctggg 4860tggggaaagt aaaaaatatg
ctggttcaag gtctaaagta aaatgataaa taatgtttgt 4920agcattaatg
aaatattttc aagaaatgtg tccgggggta gcattggcta tgctgacgag
4980gcctttggta actcagaaag ctcttggccc cgatggcgac ttgcccttgc
actttcttta 5040tcaggctctg agctcacacg gagcctctgg catttccctg
ctgtcttggg agaaaggaaa 5100ctggttgtgg cggcagggtg tggaatctgc
tgctggaacc aggctggaag cccacctggt 5160agtgaacagg gcccagcggg
gcaggctagg agtgttgtgg tctatgggtt tgtgtcctgg 5220agaatgttca
agaatgtctt cttggctgct ttggtgctga gctctgttat ctcacagcac
5280gtcctgaagg ctaacccagg tggggaggat gctgacacca gctccaggtg
gagttggtga 5340gaaatctgtc ttaacttgga gatgcagggg caacctgtga
ccctttgagg caagagccct 5400gcacccagct gtcccgtgca gccgtgggca
gggggctgca catggagggg caggcgggcc 5460agttcagggc cagttcagtg
ccctgtaagg gcccttcagc ctcctgtcct ctgtgcggct 5520gggcgccagc
accagggagt ttctatggca accttagtga ttattaagga acattgtcag
5580ttttatgaac atatgctcaa atgaaattct actttaggag gaaaggattg
gaacagcatg 5640ttgcaaggct gttaattaac agagagacct tattggatgg
agatcacatc tgttaaatag 5700aatacctcaa ctctacgttg ttttcttgga
gataaataat agtttcaagt ttttgtttgt 5760ttgttttacc taattacctg
aaagcaaata ccaaaggctg atgtctgtat atggggcaaa 5820gggtcagtat
atttttcagt gttttttttt cttttacaag ctattttgca tttaaagtga
5880acattgtaaa tgtttgtaat aaatgatttt taaaaataca 592010449PRTMacaca
fascicularis 10Leu Ala Pro Gly Gly Cys Pro Ala Gln Glu Val Ala Arg
Gly Val Leu 1 5 10 15Thr Ser Leu Pro Gly Asp Ser Val Thr Leu Thr
Cys Pro Gly Gly Glu 20 25 30Pro Glu Asp Asn Ala Thr Val His Trp Val
Leu Arg Lys Pro Ala Glu 35 40 45Gly Ser His Leu Ser Arg Trp Ala Gly
Val Gly Arg Arg Leu Leu Leu 50 55 60Arg Ser Val Gln Leu His Asp Ser
Gly Asn Tyr Ser Cys Tyr Arg Ala65 70 75 80Gly Arg Pro Ala Ala Thr
Val His Leu Leu Val Asp Val Pro Pro Glu 85 90 95Glu Pro Gln Leu Ser
Cys Phe Arg Lys Ser Pro Leu Ser Asn Val Val 100 105 110Cys Glu Trp
Gly Pro Arg Ser Thr Pro Ser Pro Thr Thr Lys Ala Val 115 120 125Leu
Leu Val Arg Lys Phe Gln Asn Ser Pro Ala Glu Asp Phe Gln Glu 130 135
140Pro Cys Gln Tyr Ser Gln Glu Ser Gln Lys Phe Ser Cys Gln Leu
Ala145 150 155 160Val Pro Glu Gly Asp Ser Ser Phe Tyr Ile Val Ser
Met Cys Val Ala 165 170 175Ser Ser Val Gly Ser Lys Leu Ser Lys Thr
Gln Thr Phe Gln Gly Cys 180 185 190Gly Ile Leu Gln Pro Asp Pro Pro
Ala Asn Ile Thr Val Thr Ala Val 195 200 205Ala Arg Asn Pro Arg Trp
Leu Ser Val Thr Trp Gln Asp Pro His Ser 210 215 220Trp Asn Ser Ser
Phe Tyr Arg Leu Arg Phe Glu Leu Arg Tyr Arg Ala225 230 235 240Glu
Arg Ser Lys Thr Phe Thr Thr Trp Met Val Lys Asp Leu Gln His 245 250
255His Cys Val Ile His Asp Ala Trp Ser Gly Leu Arg His Val Val Gln
260 265 270Leu Arg Ala Gln Glu Glu Phe Gly Gln Gly Glu Trp Ser Glu
Trp Ser 275 280 285Pro Glu Ala Met Gly Thr Pro Trp Thr Glu Ser Arg
Ser Pro Pro Ala 290 295 300Glu Asn Glu Val Ser Thr Pro Thr Gln Ala
Pro Thr Thr Asn Lys Asp305 310 315 320Asp Asp Asn Ile Leu Ser Gly
Asp Ser Ala Asn Ala Thr Ser Leu Pro 325 330 335Val Gln Asp Ser Ser
Ser Val Pro Leu Pro Thr Phe Leu Val Ala Gly 340 345 350Gly Ser Leu
Ala Phe Gly Thr Leu Leu Cys Ile Ala Ile Val Leu Arg 355 360 365Phe
Lys Lys Thr Trp Lys Leu Arg Ala Leu Lys Glu Gly Lys Thr Ser 370 375
380Met His Pro Pro Tyr Ser Leu Gly Gln Leu Val Pro Glu Arg Pro
Arg385 390 395 400Pro Thr Pro Val Leu Val Pro Leu Ile Ser Pro Pro
Val Ser Pro Ser 405 410 415Ser Leu Gly Ser Asp Asn Thr Ser Ser His
Asn Arg Pro Asp Ala Arg 420 425 430Asp Pro Arg Ser Pro Tyr Asp Ile
Ser Asn Thr Asp Tyr Phe Phe Pro 435 440 445Arg 11468PRTHomo sapiens
11Met Leu Ala Val Gly Cys Ala Leu Leu Ala Ala Leu Leu Ala Ala Pro 1
5 10 15Gly Ala Ala Leu Ala Pro Arg Arg Cys Pro Ala Gln Glu Val Ala
Arg 20 25 30Gly Val Leu Thr Ser Leu Pro Gly Asp Ser Val Thr Leu Thr
Cys Pro 35 40 45Gly Val Glu Pro Glu Asp Asn Ala Thr Val His Trp Val
Leu Arg Lys 50 55 60Pro Ala Ala Gly Ser His Pro Ser Arg Trp Ala Gly
Met Gly Arg Arg65 70 75 80Leu Leu Leu Arg Ser Val Gln Leu His Asp
Ser Gly Asn Tyr Ser Cys 85 90 95Tyr Arg Ala Gly Arg Pro Ala Gly Thr
Val His Leu Leu Val Asp Val 100 105 110Pro Pro Glu Glu Pro Gln Leu
Ser Cys Phe Arg Lys Ser Pro Leu Ser 115 120 125Asn Val Val Cys Glu
Trp Gly Pro Arg Ser Thr Pro Ser Leu Thr Thr 130 135 140Lys Ala Val
Leu Leu Val Arg Lys Phe Gln Asn Ser Pro Ala Glu Asp145 150 155
160Phe Gln Glu Pro Cys Gln Tyr Ser Gln Glu Ser Gln Lys Phe Ser Cys
165 170 175Gln Leu Ala Val Pro Glu Gly Asp Ser Ser Phe Tyr Ile Val
Ser Met 180 185 190Cys Val Ala Ser Ser Val Gly Ser Lys Phe Ser Lys
Thr Gln Thr Phe 195 200 205Gln Gly Cys Gly Ile Leu Gln Pro Asp Pro
Pro Ala Asn Ile Thr Val 210 215 220Thr Ala Val Ala Arg Asn Pro Arg
Trp Leu Ser Val Thr Trp Gln Asp225 230 235 240Pro His Ser Trp Asn
Ser Ser Phe Tyr Arg Leu Arg Phe Glu Leu Arg 245 250 255Tyr Arg Ala
Glu Arg Ser Lys Thr Phe Thr Thr Trp Met Val Lys Asp 260 265 270Leu
Gln His His Cys Val Ile His Asp Ala Trp Ser Gly Leu Arg His 275 280
285Val Val Gln Leu Arg Ala Gln Glu Glu Phe Gly Gln Gly Glu Trp Ser
290 295 300Glu Trp Ser Pro Glu Ala Met Gly Thr Pro Trp Thr Glu Ser
Arg Ser305 310 315 320Pro Pro Ala Glu Asn Glu Val Ser Thr Pro Met
Gln Ala Leu Thr Thr 325 330 335Asn Lys Asp Asp Asp Asn Ile Leu Phe
Arg Asp Ser Ala Asn Ala Thr 340 345 350Ser Leu Pro Val Gln Asp Ser
Ser Ser Val Pro Leu Pro Thr Phe Leu 355 360 365Val Ala Gly Gly Ser
Leu Ala Phe Gly Thr Leu Leu Cys Ile Ala Ile 370 375 380Val Leu Arg
Phe Lys Lys Thr Trp Lys Leu Arg Ala Leu Lys Glu Gly385 390 395
400Lys Thr Ser Met His Pro Pro Tyr Ser Leu Gly Gln Leu Val Pro Glu
405 410 415Arg Pro Arg Pro Thr Pro Val Leu Val Pro Leu Ile Ser Pro
Pro Val 420 425 430Ser Pro Ser Ser Leu Gly Ser Asp Asn Thr Ser Ser
His Asn Arg Pro 435 440 445Asp Ala Arg Asp Pro Arg Ser Pro Tyr Asp
Ile Ser Asn Thr Asp Tyr 450 455 460Phe Phe Pro Arg465121718DNAHomo
sapiens 12tgagtcatgt gcgagtggga agtcgcactg acactgagcc gggccagagg
gagaggagcc 60gagcgcggcg cggggccgag ggactcgcag tgtgtgtaga gagccgggct
cctgcggatg 120ggggctgccc ccggggcctg agcccgcctg cccgcccacc
gccccgcccc gcccctgcca 180cccctgccgc ccggttccca ttagcctgtc
cgcctctgcg ggaccatgga gtggtagccg 240aggaggaagc atgctggccg
tcggctgcgc gctgctggct gccctgctgg ccgcgccggg 300agcggcgctg
gccccaaggc gctgccctgc gcaggaggtg gcgagaggcg tgctgaccag
360tctgccagga gacagcgtga ctctgacctg cccgggggta gagccggaag
acaatgccac 420tgttcactgg gtgctcagga agccggctgc aggctcccac
cccagcagat gggctggcat 480gggaaggagg ctgctgctga ggtcggtgca
gctccacgac tctggaaact attcatgcta 540ccgggccggc cgcccagctg
ggactgtgca cttgctggtg gatgttcccc ccgaggagcc 600ccagctctcc
tgcttccgga agagccccct cagcaatgtt gtttgtgagt ggggtcctcg
660gagcacccca tccctgacga caaaggctgt gctcttggtg aggaagtttc
agaacagtcc 720ggccgaagac ttccaggagc cgtgccagta ttcccaggag
tcccagaagt tctcctgcca 780gttagcagtc ccggagggag acagctcttt
ctacatagtg tccatgtgcg tcgccagtag 840tgtcgggagc aagttcagca
aaactcaaac ctttcagggt tgtggaatct tgcagcctga 900tccgcctgcc
aacatcacag tcactgccgt ggccagaaac ccccgctggc tcagtgtcac
960ctggcaagac ccccactcct ggaactcatc tttctacaga ctacggtttg
agctcagata 1020tcgggctgaa cggtcaaaga cattcacaac atggatggtc
aaggacctcc agcatcactg 1080tgtcatccac gacgcctgga gcggcctgag
gcacgtggtg cagcttcgtg cccaggagga 1140gttcgggcaa ggcgagtgga
gcgagtggag cccggaggcc atgggcacgc cttggacaga 1200atccaggagt
cctccagctg agaacgaggt gtccaccccc atgcaggcac ttactactaa
1260taaagacgat gataatattc tcttcagaga ttctgcaaat gcgacaagcc
tcccagtgca 1320agattcttct tcagtaccac tgcccacatt cctggttgct
ggagggagcc tggccttcgg 1380aacgctcctc tgcattgcca ttgttctgag
gttcaagaag acgtggaagc tgcgggctct 1440gaaggaaggc aagacaagca
tgcatccgcc gtactctttg gggcagctgg tcccggagag 1500gcctcgaccc
accccagtgc ttgttcctct catctcccca ccggtgtccc ccagcagcct
1560ggggtctgac aatacctcga gccacaaccg accagatgcc agggacccac
ggagccctta 1620tgacatcagc aatacagact acttcttccc cagatagctg
gctgggtggc accagcagcc 1680tggaccctgt ggatgataaa acacaaacgg gctcagca
171813456DNAArtificial SequenceConsensus sequence of human gp80 and
cyno gp80. 13Met Leu Ala Val Gly Cys Ala Leu Leu Ala Ala Leu Leu
Ala Pro Gly 1 5 10 15Ala Ala Leu Ala Pro Cys Pro Ala Gln Glu Val
Ala Arg Gly Val Leu 20 25 30Thr Ser Leu Pro Gly Asp Ser Val Thr Leu
Thr Cys Pro Gly Glu Pro 35 40 45Glu Asp Asn Ala Thr Val His Trp Val
Leu Arg Lys Pro Ala Gly Ser 50 55 60His Ser Arg Trp Ala Gly Met Gly
Arg Arg Leu Leu Leu Arg Ser Val65 70 75 80Gln Leu His Asp Ser Gly
Asn Tyr Ser Cys Tyr Arg Ala Gly Arg Pro 85 90 95Ala Ala Thr Val His
Leu Leu Val Asp Val Pro Pro Glu Glu Pro Gln 100 105 110Leu Ser Cys
Phe Arg Lys Ser Pro Leu Ser Asn Val Val Cys Glu Trp 115 120 125Gly
Pro Arg Ser Thr Pro Ser Thr Thr Lys Ala Val Leu Leu Val Arg 130 135
140Lys Phe Gln Asn Ser Pro Ala Glu Asp Phe Gln Glu Pro Cys Gln
Tyr145 150 155 160Ser Gln Glu Ser Gln Lys Phe Ser Cys Gln Leu Ala
Val Pro Glu Gly 165 170 175Asp Ser Ser Phe Tyr Ile Val Ser Met Cys
Val Ala Ser Ser Val Gly 180 185 190Ser Lys Ser Lys Thr Gln Thr Phe
Gln Gly Cys Gly Ile Leu Gln Pro 195 200 205Asp Pro Pro Ala Asn Ile
Thr Val Thr Ala Val Ala Arg Asn Pro Arg 210 215 220Trp Leu Ser Val
Thr Trp Gln Asp Pro His Ser Trp Asn Ser Ser Phe225 230 235 240Tyr
Arg Leu Arg Phe Glu Leu Arg Tyr Arg Ala Glu Arg Ser Lys Thr 245 250
255Phe Thr Thr Trp Met Val Lys Asp Leu Gln His His Cys Val Ile His
260 265 270Asp Ala Trp Ser Gly Leu Arg His Val Val Gln Leu Arg Ala
Gln Glu 275 280 285Glu Phe Gly Gln Gly Glu Trp Ser Glu Trp Ser Pro
Glu Ala Met Gly 290 295 300Thr Pro Trp Thr Glu Ser Arg Ser Pro Pro
Ala Glu Asn Glu Val Ser305 310 315 320Thr Pro Gln Ala Thr Thr Asn
Lys Asp Asp Asp Asn Ile Leu Asp Ser 325 330 335Ala Asn Ala Thr Ser
Leu Pro Val Gln Asp Ser Ser Ser Val Pro Leu 340 345 350Pro Thr Phe
Leu Val Ala Gly Gly Ser Leu Ala Phe Gly Thr Leu Leu 355 360 365Cys
Ile Ala Ile Val Leu Arg Phe Lys
Lys Thr Trp Lys Leu Arg Ala 370 375 380Leu Lys Glu Gly Lys Thr Ser
Met His Pro Pro Tyr Ser Leu Gly Gln385 390 395 400Leu Val Pro Glu
Arg Pro Arg Pro Thr Pro Val Leu Val Pro Leu Ile 405 410 415Ser Pro
Pro Val Ser Pro Ser Ser Leu Gly Ser Asp Asn Thr Ser Ser 420 425
430His Asn Arg Pro Asp Ala Arg Asp Pro Arg Ser Pro Tyr Asp Ile Ser
435 440 445Asn Thr Asp Tyr Phe Phe Pro Arg 450 455
* * * * *
References